Inhibitors of Flaviviridae viruses

ABSTRACT

Provided are compounds of Formula I: and pharmaceutically acceptable salts and esters thereof. The compounds, compositions, and methods provided are useful for the treatment of Flaviviridae virus infections, particularly hepatitis C infections.

FIELD OF THE INVENTION

The present application includes novel inhibitors of Flaviviridaeviruses, compositions containing such compounds, therapeutic methodsthat include the administration of such compounds.

BACKGROUND OF THE INVENTION

Viruses comprising the Flaviviridae family include at least threedistinguishable genera including pestiviruses, flaviviruses, andhepaciviruses (Calisher, et al., J. Gen. Virol., 1993, 70, 37-43). Whilepestiviruses cause many economically important animal diseases such asbovine viral diarrhea virus (BVDV), classical swine fever virus (CSFV,hog cholera) and border disease of sheep (BDV), their importance inhuman disease is less well characterized (Moennig, V., et al., Adv. Vir.Res. 1992, 48, 53-98). Flaviviruses are responsible for important humandiseases such as dengue fever and yellow fever while hepaciviruses causehepatitis C virus infections in humans. Other important viral infectionscaused by the Flaviviridae family include West Nile virus (WNV) Japaneseencephalitis virus (JEV), tick-borne encephalitis virus, Junjin virus,Murray Valley encephalitis, St Louis enchaplitis, Omsk hemorrhagic fevervirus and Zika virus.

The hepatitis C virus (HCV) is the leading cause of chronic liverdisease worldwide (Boyer, N. et al. J Hepatol. 32:98-112, 2000) so asignificant focus of current antiviral research is directed toward thedevelopment of improved methods of treatment of chronic HCV infectionsin humans (Di Besceglie, A. M. and Bacon, B. R., Scientific American,Oct.: 80-85, (1999); Gordon, C. P., et al., J. Med. Chem. 2005, 48,1-20; Maradpour, D., et al., Nat. Rev. Micro. 2007, 5(6), 453-463). Anumber of HCV treatments are reviewed by Dymock et al. in AntiviralChemistry & Chemotherapy, 11:2; 79-95 (2000). Virologic cures ofpatients with chronic HCV infection are difficult to achieve because ofthe prodigious amount of daily virus production in chronically infectedpatients and the high spontaneous mutability of HCV virus (Neumann, etal., Science 1998, 282, 103-7; Fukimoto, et al., Hepatology, 1996, 24,1351-4; Domingo, et al., Gene, 1985, 40, 1-8; Martell, et al., J. Viral.1992, 66, 3225-9.

Currently, there are primarily two antiviral compounds, ribavirin, anucleoside analog, and interferon-alpha (α) (IFN), that are used for thetreatment of chronic HCV infections in humans. Ribavirin alone is noteffective in reducing viral RNA levels, has significant toxicity, and isknown to induce anemia. The combination of IFN and ribavirin has beenreported to be effective in the management of chronic hepatitis C(Scott, L. J., et al. Drugs 2002, 62, 507-556) but less than half thepatients given this treatment show a persistent benefit.

Combined, infections from the Flaviviridae virus family causesignificant mortality, morbidity and economic losses throughout theworld. Alkynyl substituted thiophenes with anti-Flaviviridae virusactivity have been disclosed by Chan, et al., WO 2008058393; Wunberg, etal., WO 2006072347; and Chan, et al., WO 2002100851; but none of theseare currently clinically approved antiviral therapeutics. Therefore,there remains a need to develop effective treatments for Flaviviridaevirus infections.

SUMMARY OF THE INVENTION

In one aspect, provided is a compound of Formula I:

or a pharmaceutically acceptable salt or ester thereof, wherein;

R¹ is -(L¹)_(m)-(L²)_(n)-X¹;

L¹ is selected from the group consisting of alkynylene and optionallysubstituted arylene;

m is 1;

when L¹ is substituted, L¹ is substituted with one or more Q⁶;

L² is selected from the group consisting of —NHC(O)— and optionallysubstituted alkylene;

n is 0 or 1;

when L² is substituted, L² is substituted with one or more Q⁶;

X¹ is selected from the group consisting of

-   -   a)—P(O)R^(x)R^(y),    -   b) —P(O)OR^(x)R^(y),    -   c)—OP(O)R^(x)R^(y) and    -   d) optionally substituted C₁₋₁₂ alkyl, optionally substituted        C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally        substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl,        optionally substituted 3-14 membered heteroaryl, optionally        substituted 3-12 membered heterocyclyl, optionally substituted        3-18 membered heteroarylalkyl and optionally substituted C₆₋₁₈        arylalkyl;

each of R^(x) and R^(y) independently is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

or R^(x) and R^(y) taken together with the atoms to which they areattached form a 3 to 10 membered heterocyclyl;

when either R^(x) or R^(y) is substituted, R^(x) or R^(y) is substitutedwith one or more Q⁶;

when X¹ is selected from d) and is substituted, X¹ is substituted withone or more Q¹;

each Q¹ individually is selected from the group consisting of halogen,oxo, oxide, —NO₂, —SR¹¹, —S(O)R¹⁰, —S(O)₂R¹⁰, —S(O)₂NR¹²R¹¹,—NR¹²C(O)R¹¹, —NR¹²C(O)NR¹¹R¹², —NR¹¹S(O)R¹⁰, —NR¹¹S(O)₂R¹⁰,—NR¹²S(O)₂NR¹¹R¹², —CR¹²(═NNR¹¹R¹²), —CR¹²(═NOR¹¹), —ONR¹²R¹¹,—ON(═CR¹²R¹¹), —NR¹²OR¹¹, —OH, —NR¹¹R¹², —C(O)OR¹², —CN, —N₃,—C(═NR¹³)NR¹¹R¹², —NR¹²C(═NR¹³)NR¹¹R¹², —NR¹²C(O)OR¹¹, —OC(O)NR¹¹R¹²,—OP(O)R¹¹R¹², —P(O)R¹¹R¹², —P(O)OR¹¹R¹², —C(O)NR¹¹R¹², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(±)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted-3-10 membered heterocyclyl;

wherein each R¹⁰, independently, is selected from the group consistingof optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

wherein each R¹¹ and R¹², independently, is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted O₂₋₁₂ alkenyl, optionally substituted O₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

or R¹¹ and R¹² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

or R¹⁰ and R¹¹ taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

each R¹³ independently is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl, optionally substituted C₈₋₁₈ arylalkyl, —CN, —C(O)R¹⁴,—CHO and —S(O)₂R¹⁴;

where each R¹⁴ individually is optionally substituted C₁₋₁₂ alkyl;

when Q¹ is substituted, Q¹ is substituted with one or more Q⁶;

R² is selected from the group consisting of optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂ cycloalkyl, optionallysubstituted C₆₋₁₄ aryl, optionally substituted 3-14 membered heteroaryl,optionally substituted 3-12 membered heterocyclyl, optionallysubstituted 3-18 membered heteroarylalkyl and optionally substitutedC₆₋₁₈ arylalkyl;

wherein, when R² is substituted, R² is substituted with one or more Q²;

where each Q² individually is selected from the group consisting ofhalogen, oxo, oxide, —NO₂, —SR²¹, —S(O)R²⁰, —S(O)₂R²⁰, —S(O)₂NR²²R²¹,—NR²²C(O)R²¹, —NR²²C(O)NR²¹R²², —NR²¹S(O)R²⁰, —NR²¹S(O)₂R²⁰,—NR²²S(O)₂NR²¹R²², —CR²²(═NNR²¹R²²), —CR²²(═NOR²¹), —ONR²²R²¹,—ON(═CR²²R²¹), —NR²²OR²¹, —OH, —NR²¹R²², —C(O)OR²², —CN, —N₃, —C(═NR²³)NR²¹R²², —NR²²C(═NR²³)N R²¹R²², —NR²²C(O)OR²¹, and —OC(O)NR²¹R²²,—OP(O)R²¹R²², —P(O)R²¹R²², —P(O)OR²¹R²², —C(O)NR²¹R²², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₈ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(±)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted 3-10 membered heterocyclyl;

wherein each R²⁰, independently, is selected from the group consistingof optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

wherein each R²¹ and R²², independently, is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

or R²¹ and R²² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

or R²⁰ and R²¹ taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

each R²³ independently is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl, optionally substituted C₆₋₁₈ arylalkyl, —CN, —C(O)R²⁴,CHO and —S(O)₂R²⁴;

where each R²⁴ individually is optionally substituted C₁₋₁₂ alkyl;

when Q² is substituted, Q² is substituted with one or more Q⁶;

R³ is -(L³)_(p)-(L⁴)_(q)-(X³);

L³ is selected from the group consisting of optionally substituted C₁₋₁₂alkylene, optionally substituted C₂₋₁₂ alkenylene, optionallysubstituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene, optionally substituted C₁₋₁₂ haloalkylene, optionallysubstituted C₆₋₁₄ arylene, optionally substituted 3-12 memberedheterocyclylene, optionally substituted 3-18 membered heteroarylalkyleneand optionally substituted C₆₋₁₈ arylalkylene;

p is 0 or 1;

when L³ is substituted, L³ is substituted with one or more Q³,

each Q³ individually is selected from the group consisting of halogen,oxo, oxide, —NO₂, —SR³¹, —S(O)R³⁰, —S(O)₂R³⁰, —S(O)₂NR³²R³¹,—NR³²C(O)R³¹, NR³²C(O)NR³¹R³², —NR³¹S(O)R³⁰, —NR³¹S(O)₂R³⁰,—NR³²S(O)₂NR³¹R³², —CR³²(═NNR³¹R³²), —CR³²(═NOR³¹), —ONR³²R³¹,—ON(═CR³²R³¹), —NR³²OR³¹, —OH, —NR³¹R³², —C(O)OR³², —CN, —N₃, —C(═NR³³)NR³¹R³², —NR³⁰C(═NR³³)N R³¹R³², —NR³⁰C(O)OR³¹, and —OC(O)NR³¹R³²,—OP(O)R³¹R³², —P(O)R³¹R³², —P(O)OR³¹R³², —C(O)NR³¹R³², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(O)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted 3-10 membered heterocyclyl;

wherein each R³⁰, independently, is selected from the group consistingof optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted O₆₋₁₈ arylalkyl;

wherein each R³¹ and R³², independently, is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C_(e-14)aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

or R³¹ and R³² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

or R³⁰ and R³¹ taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

each R³³ independently is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl, optionally substituted C₆₋₁₈ aryfalkyl, —CN, —C(O)R³⁴,CHO and —S(O)₂R³⁴;

where each R³⁴ individually is optionally substituted C₁₋₁₂ alkyl;

when Q³ is substituted, Q³ is substituted with one or more Q⁶;

L⁴ is selected from the group consisting of —NH— and optionallysubstituted C₁₋₁₂ alkylene;

q is 0, 1 or 2;

when L⁴ is substituted, L⁴ is substituted with one or more Q⁶;

X³ is selected from the group consisting of:

-   -   e) —P(O)R^(x)R^(y),    -   f) —P(O)OR^(x)R^(y),    -   g) —OP(O)R^(x)R^(y),    -   h) —P(O)(OR^(x))(OR^(y)),    -   i)

-   -   wherein each A is a 4 to 6 membered ring, including the depicted        P atom,    -   j)

and

-   -   k) optionally substituted C₁₋₁₂ alkyl, optionally substituted        C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally        substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl,        optionally substituted 3-14 membered heteroaryl, optionally        substituted 3-12 membered heterocyclyl, optionally substituted        3-18 membered heteroarylalkyl and optionally substituted C₆₋₁₈        arylalkyl;

each of R^(x) and R^(y) individually is as defined above;

when X³ is selected from k) and is substituted, X³ is substituted withone or more Q⁴;

each Q⁴ individually is selected from the group consisting of halogen,oxo, oxide, —NO₂, —SR⁴¹, —S(O)R⁴⁰, —S(O)₂R⁴⁰, —S(O)₂NR⁴²R⁴¹,S(O)₂NR⁴²R⁴¹, —NR⁴²C(O)R⁴¹, —NR⁴²C(O)NR⁴¹R⁴², —NR⁴¹S(O)R⁴⁰,—NR⁴¹S(O)₂R⁴⁰, —NR⁴²S(O)₂NR⁴¹R⁴², —CR⁴²(═NNR⁴¹R⁴²), —CR⁴²(═NOR⁴¹),—ONR⁴²R⁴¹, —ON(═CR⁴²R⁴¹), —NR⁴²OR⁴¹, —OH, —NR⁴¹R⁴², —C(O)OR⁴², —CN, —N₃,—C(═NR⁴³)NR⁴¹R⁴², —NR⁴²C(═NR⁴³)NR⁴¹R⁴², —NR⁴²C(O)OR⁴¹, and—OC(O)NR⁴¹R⁴², —OP(O)R⁴¹R⁴², —P(O)R⁴¹R⁴², —P(O)OR⁴¹R⁴², —C(O)NR⁴¹R⁴²,optionally substituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl,optionally substituted C₂₋₆ alkynyl, optionally substituted C₃₋₆cycloalkyl, optionally substituted C₆₋₁₂ arylalkyl, optionallysubstituted C₆₋₁₂ aryl, optionally substituted 3-14 membered heteroaryl,optionally substituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆alkenyloxy, optionally substituted C₂₋₆ alkynyloxy, optionallysubstituted C₃₋₆ cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy,optionally substituted 3-14 membered heteroaryloxy, optionallysubstituted 4-12 membered heterocyclyloxy, optionally substituted—C(O)C₁₋₆ alkyl, optionally substituted —C(O)C₂₋₆ alkenyl, optionallysubstituted —C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆cycloalkyl, optionally substituted —C(O)C₆₋₁₂ aryl, optionallysubstituted —C(±)-3-14 membered heteroaryl, optionally substituted—C(O)C₆₋₁₂ arylalkyl and optionally substituted-3-10 memberedheterocyclyl;

wherein each R⁴⁰, independently, is selected from the group consistingof optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

wherein each R⁴¹ and R⁴², independently, is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;

or R⁴¹ and R⁴² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

or R⁴⁰ and R⁴¹ taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

each R⁴³ independently is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl, optionally substituted C₆₋₁₆ arylalkyl, —CN, —C(O)R⁴⁴,CHO and —S(O)₂R⁴⁴;

where each R⁴⁴ individually is optionally substituted C₁₋₁₂ alkyl;

when Q⁴ is substituted, Q⁴ is substituted with one or more Q⁶; and

each Q⁶ individually is selected from the group consisting of halogen,oxo, oxide, —NO₂, —N(═O), —SR⁶¹, —S(O)R⁶⁰, —S(O)₂R⁶⁰, —S(O)₂NR⁶²R⁶¹,—NR⁶²C(O)R⁶¹, —NR⁶²C(O)NR⁶¹R⁶², —NR⁶¹S(O)R⁶⁰, —NR⁶¹S(O)₂R⁶⁰,—NR⁶²S(O)₂NR⁶¹R⁶², —CR⁶²(═NNR⁶¹R⁶²), —CR⁶²(═NOR⁶¹), —ONR⁶²R⁶¹,—ON(═CR⁶²R⁶¹), —NR⁶²OR⁶¹, —OH, —NR⁶¹R⁶², C(O)OR⁶², —CN, —N₃, —C(═NR⁶³)NR⁶¹R⁶², —NR⁶²C(═NR⁶³)N R⁶¹R⁶², —NR⁶²C(O)OR⁶¹, and —OC(O)NR⁶¹R⁶²,—OP(O)R⁶¹R⁶², P(O)R⁶¹R⁶², —P(O)OR⁶¹R⁶², —P(O)(OR⁶¹)OR⁶², —C(O)NR⁶¹R⁶²,C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, C₆₋₁₂arylalkyl, C₆₋₁₂ aryl, 3-14 membered heteroaryl, C₁₋₆ alkyloxy, C₂₋₆alkenyloxy, C₂₋₆ alkynyloxy, C₃₋₆ cycloalkyloxy, C₆₋₁₂ aryloxy, 3-14membered heteroaryloxy, 4-12 membered heterocyclyloxy, —C(O)C₁₋₆ alkyl,—C(O)C₂₋₆ alkenyl, —C(O)C₂₋₆ alkynyl, —C(O)C₃₋₆ cycloalkyl, —C(O)C₁₋₆haloalkyl, —C(O)C₆₋₁₂ aryl, —C(O)— 3-14 membered heteroaryl, —C(O)C₆₋₁₂arylalkyl and 3-10 membered heterocyclyl;

wherein each R⁶⁰, independently, is selected from the group consistingof C₁₋₁₂ alkyl, O₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂ cycloalkyl, C₁₋₁₂haloalkyl, C₆₋₁₄ aryl, 3-14 membered heteroaryl, 3-12 memberedheterocyclyl, 3-18 membered heteroarylalkyl and C₆₋₁₈ arylalkyl;

wherein each R⁶¹ and R⁶², independently, is selected from the groupconsisting of H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂cycloalkyl, C₁₋₁₂ haloalkyl, C₆₋₁₄ aryl, 3-14 membered heteroaryl, 3-12membered heterocyclyl, 3-18 membered heteroarylalkyl and C₆₋₁₈arylalkyl;

or R⁶¹ and R⁶² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

or R⁶⁰ and R⁶¹ taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl;

each R⁶³ independently is selected from the group consisting of H, C₁₋₁₂alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂ cycloalkyl, C₆₋₁₄ aryl, 3-14membered heteroaryl, 3-12 membered heterocyclyl, 3-18 memberedheteroarylalkyl, C₆₋₁₈ arylalkyl, —CN, —C(O)R⁶⁴, —CHO and —S(O)₂R⁶⁴;

where each R⁶⁴ individually is C₁₋₁₂ alkyl;

provided, however, that when X¹ is selected from d), then X³ is selectedfrom the group consisting of e), f), g), h), i) and j).

In another aspect, a method for treating Flaviviridae viral infectionsis provided comprising administering a therapeutically effective amountof a compound of Formula I to a mammal in need thereof. The compound ofFormula I is administered to a human subject in need thereof, such as ahuman being who is infected with viruses of the Flaviviridae family. Inanother embodiment, the compound of Formula I is administered to a humansubject in need thereof, such as a human being who is infected with aHCV virus. In one embodiment, the treatment results in the reduction ofone or more of the in viral loads or clearance of viral RNA in apatient.

In another embodiment, provided is a method of treating and/orpreventing a disease caused by a viral infection wherein the viralinfection is caused by a virus selected from the group consisting ofdengue virus, yellow fever virus, West Nile virus, Japanese encephalitisvirus, tick-borne encephalitis virus, Junjin virus, Murray Valleyencephalitis virus, St Louis encephalitis virus, Omsk hemorrhagic fevervirus, bovine viral disarrhea virus, Zika virus and Hepatitis C virus;by administering to a subject in need thereof a therapeuticallyeffective amount of a compound of Formula I, or a pharmaceuticallyacceptable salt or ester thereof.

In another aspect, provided is the use of a compound of Formula I forthe manufacture of a medicament for the treatment of Flaviviridae viralinfections. In another aspect, provided is a compound of Formula I foruse in treating a Flaviviridae viral infection. In one embodiment, theFlaviviridae viral infection is acute or chronic HCV infection. In oneembodiment of each aspect of use and compound, the treatment results inthe reduction of one or more of the viral loads or clearance of RNA inthe patient.

In another aspect, provided is a method for treating or preventing HCVcomprising administering an effective amount of a compound of Formula Ito a patient in need thereof. In another aspect, provided is the use ofa compound of the present invention for the manufacture of a medicamentfor the treatment or prevention of HCV.

In another aspect, provided is a use of a compound of Formula I for thetreatment of a Flaviviridae viral infection or a Hepatitis C virusinfection.

In another aspect, provided is a pharmaceutical composition comprising acompound of Formula I or a pharmaceutically acceptable salt or esterthereof and one or more pharmaceutically acceptable carriers orexcipients. The pharmaceutical composition of Formula I may furthercomprise one or more additional therapeutic agents. The one or moreadditional therapeutic agent may be, without limitation, selected from:interferons, ribavirin or its analogs, HCV NS3 protease inhibitors, NS5ainhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants,mevalonate decarboxylase antagonists, antagonists of therenin-angiotensin system, other anti-fibrotic agents, endothelinantagonists, nucleoside or nucleotide inhibitors of HCV NS5B polymerase,non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors,TLR-7 agonists, cyclophillin inhibitors, HCV IRES inhibitors,pharmacokinetic enhancers and other drugs for treating HCV; or mixturesthereof.

In another aspect, provided is a method for the treatment or preventionof the symptoms or effects of an HCV infection in an infected animalwhich comprises administering to, i.e. treating, said animal with apharmaceutical combination composition or formulation comprising aneffective amount of a Formula I compound, and a second compound havinganti-HCV properties.

In another embodiment, provided are compounds of Formula I andpharmaceutically acceptable salts and esters thereof and all racemates,enantiomers, diastereomers, tautomers, polymorphs, pseudopolymorphs andamorphous forms thereof.

In another aspect, provided are processes and novel intermediatesdisclosed herein which are useful for preparing Formula I compounds.

In other aspects, novel methods for synthesis, analysis, separation,isolation, purification, characterization, and testing of the compoundsof Formula I are provided.

The present invention includes combinations of aspects and embodiments,as well as preferences, as herein described throughout the presentspecification.

DETAILED DESCRIPTION

Reference will now be made in detail to certain embodiments of theinvention, examples of which are illustrated in the accompanyingstructures and formulas. While the invention will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the invention to those embodiments. Onthe contrary, the invention is intended to cover all alternatives,modifications, and equivalents, which may be included within the scopeof the present invention as defined herein.

Each document referenced herein is incorporated by reference in itsentirety for all purposes.

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isalkynylene and n is 0. In another aspect of this embodiment, L¹ isethynylene. In another aspect of this embodiment, X¹ is optionallysubstituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂ alkenyl,optionally substituted C₂₋₁₂ alkynyl or optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, X¹ is optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X¹ isoptionally substituted C₃-C₇ secondary or tertiary alkyl. In anotheraspect of this embodiment, X¹ is optionally substituted C₃-C₅cycloalkyl. In another aspect of this embodiment, X¹ is prop-2-yl(isopropyl) or 2-methylprop-2-yl (1-butyl).

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isalkynylene and n is 1. In another aspect of this embodiment, L¹ isethynylene. In another aspect of this embodiment, L² is optionallysubstituted C₁₋₁₂ alkylene. In another aspect of this embodiment, L² isoptionally substituted C₅-C₇ alkylene. In another aspect of thisembodiment, X¹ is —P(O)R^(x)R^(y). In another aspect of this embodiment,X¹ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X¹ is —P(O)(CH₃)₂. In another aspect of this embodiment, X¹is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X¹ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X¹ is —P(O)(OCH₃)(CH₃). In another aspect of thisembodiment, X¹ is —OP(O)R^(x)R^(y). In another aspect of thisembodiment, X¹ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X¹ is —OP(O)(CH₃)₂.

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isoptionally substituted arylene and n is 0. In another aspect of thisembodiment, L¹ is phenylene. In another aspect of this embodiment, X¹ isoptionally substituted 3-14 membered heteroaryl. In another aspect ofthis embodiment, X¹ is a 5-10 membered heteroaryl comprising one tothree nitrogen atoms.

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isoptionally substituted arylene and n is 1. In another aspect of thisembodiment, t: is phenylene. In another aspect of this embodiment, L² isoptionally substituted C₁₋₁₂ alkylene. In another aspect of thisembodiment, L² is optionally substituted C₅-C₇ alkylene. In anotheraspect of this embodiment, L² is methylene. In another aspect of thisembodiment, L² is —NHC(O)—. In another aspect of this embodiment, X¹ is—P(O)R^(x)R^(y). In another aspect of this embodiment, X¹ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X¹ is—P(O)(CH₃)₂. In another aspect of this embodiment, X¹ is—P(O)OR^(x)R^(y). In another aspect of this embodiment, X¹ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X¹ is—P(O)(OR^(x))(CH₃). In another aspect of this embodiment, X¹ is—P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl. In another aspectof this embodiment, X¹ is —OP(O)R^(x)R^(y). In another aspect of thisembodiment, X¹ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X¹ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X¹ is optionally substituted 3-14 membered heteroaryl. Inanother aspect of this embodiment, X¹ is thiazolyl.

In another embodiment of Formula I, R² is optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂ cycloalkyl, or optionallysubstituted C₆₋₁₈ arylalkyl. In another aspect of this embodiment, R² isoptionally substituted C₃₋₁₂ cycloalkyl. In another aspect of thisembodiment, R² is optionally substituted methylcyclohexyl. In anotheraspect of this embodiment, R² is optionally substitutedmethylcyclohexenyl. In another aspect of this embodiment, R² isoptionally substituted 4-methylcyclohexyl. In a preferred aspect of thisembodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In one embodiment of Formula I, R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein pand q are each 0. In another aspect of this embodiment X³ is optionallysubstituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂ alkenyl,optionally substituted C₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionally substituted3-14 membered heteroaryl, optionally substituted 3-12 memberedheterocyclyl, optionally substituted 3-18 membered heteroarylalkyl oroptionally substituted C₆₋₁₈ arylalkyl. In another aspect of thisembodiment, X³ is optionally substituted C₃₋₁₂ cycloalkyl. In anotheraspect of this embodiment, X³ is optionally substituted cyclohexyl. Inanother aspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl.

In one embodiment of Formula I, R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein pis 1 and q is 0. In another aspect of this embodiment, L³ is optionallysubstituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂ alkenylene,optionally substituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene or optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene. In another aspect of this embodiment, L³ is optionallysubstituted ethylene. In another aspect of this embodiment, L³ isoptionally substituted C₃₋₁₂ cycloalkylene. In another aspect of thisembodiment, L³ is optionally substituted C₄₋₆ cycloalkylene. In anotheraspect of this embodiment, L³ is optionally substituted cyclohexylene.In another aspect of this embodiment, L³ is optionally substituted 3-12membered heterocyclylene. In another aspect of this embodiment, L³ isoptionally substituted 5-6 membered N-containing heterocyclene. Inanother aspect of this embodiment, L³ is piperidinylene. In anotheraspect of this embodiment, X³ is —P(O)R^(x)R^(y). In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) ia independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is —P(O)(OR^(x))(OR^(y)). In another aspect of thisembodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl.

In one embodiment of Formula I, R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein pis 1 and q is 1. In another aspect of this embodiment, L³ is optionallysubstituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂ alkenylene,optionally substituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene or optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted C₃₋₁₂cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted C₄₋₆ cycloalkylene. In another aspect of this embodiment, L³is optionally substituted cyclohexylene. In another aspect of thisembodiment, L⁴ is —NH—. In another aspect of this embodiment, L⁴ isoptionally substituted C₁₋₁₂ alkylene. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y). In another aspect of this embodiment,X³ is —P(O)R^(x)R^(Y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is —P(O)(OR^(x))(OR^(y)). In another aspect of thisembodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl.

In one embodiment of Formula I, R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein pis 1 and q is 2. In another aspect of this embodiment, L³ is optionallysubstituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂ alkenylene,optionally substituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene or optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted C₃₋₁₂cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted C₄₋₆ cycloalkylene. In another aspect of this embodiment, L³is optionally substituted cyclohexylene. In another aspect of thisembodiment, L⁴ is —NH—. In another aspect of this embodiment, L⁴ isoptionally substituted C₁₋₁₂ alkylene. In another aspect of thisembodiment, one of L⁴ is —NH— and the other L⁴ is optionally substitutedC₁₋₁₂ alkylene. In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently ethyl ormethyl. In another aspect of this embodiment, X³ is —P(O)OR^(x)R^(y). Inanother aspect of this embodiment, X³ is —P(O)OR^(x)R^(y) wherein eachR^(x) and R^(y) is independently optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃). In anotheraspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) ia Hor C₁₋₆ alkyl. In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —OP(O)(CH₃)₂. In another aspect of this embodiment, X³is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlymethyl, ethyl, or optionally substituted phenyl. In another aspect ofthis embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is —P(O)(OR^(x))(OR^(y)). In another aspect of this embodiment, X³ is—P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) is independentlymethyl or ethyl. In another aspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl.

In another embodiment of Formula I, R¹ is -(L¹)_(m)(L²)_(n)-X¹, L¹ isalkynylene, n is 0, and R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein p and qare each 0. In another aspect of this embodiment, L is ethynylene. Inanother aspect of this embodiment, X¹ is optionally substituted C₁₋₁₂alkyl, optionally substituted O₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl or optionally substituted C₃₋₁₂ cycloalkyl. In anotheraspect of this embodiment, X¹ is optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X¹ is optionally substituted C₃-C₇secondary or tertiary alkyl. In another aspect of this embodiment, X¹ isoptionally substituted C₃-C₅ cycloalkyl. In another aspect of thisembodiment, X¹ is prop-2-yl (isopropyl) or 2-methylprop-2-yl(t-butyl).

In another aspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isalkynylene, n is 0 and R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein p is 1 andq is 0. In another aspect of this embodiment, L¹ is ethynylene. Inanother aspect of this embodiment, X¹ is optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl or optionally substituted C₃₋₁₂ cycloalkyl. In anotheraspect of this embodiment, X¹ is optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X′ is optionally substituted C₃-C₇secondary or tertiary alkyl. In another aspect of this embodiment, X¹ isoptionally substituted C₃-C₅ cycloalkyl. In another aspect of thisembodiment, X¹ is prop-2-yl(isopropyl) or 2-methylprop-2-yl (t-butyl).In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene, optionally substituted C₂₋₁₂ alkenylene, optionallysubstituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂ cycloalkyleneor optionally substituted 3-12 membered heterocyclylene. In anotheraspect of this embodiment, L³ is optionally substituted C₁₋₁₂ alkylene.In another aspect of this embodiment, L³ is optionally substitutedethylene. In another aspect of this embodiment, L³ is optionallysubstituted C₃₋₁₂ cycloalkylene. In another aspect of this embodiment,L³ is optionally substituted C₄₋₆ cycloalkylene. In another aspect ofthis embodiment, L³ is optionally substituted cyclohexylene. In anotheraspect of this embodiment, L³ is optionally substituted 3-12 memberedheterocyclylene. In another aspect of this embodiment, L³ is optionallysubstituted 5-6 membered N-containing heterocyclene. In another aspectof this embodiment, L³ is piperidinylene. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y). In another aspect of this embodiment,X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is —P(O)(OR^(x))(OR^(y)), In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isalkynylene, n is 0, and R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein p is 1 andq is 1. In another aspect of this embodiment, L¹ is ethynylene. Inanother aspect of this embodiment, X¹ is optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl or optionally substituted O₃₋₁₂ cycloalkyl. In anotheraspect of this embodiment, X¹ is optimally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X¹ is optionally substituted C₃-C₇secondary or tertiary alkyl. In another aspect of this embodiment, X¹ isoptionally substituted C₃-C₅ cycloalkyl. In another aspect of thisembodiment, X¹ is prop-2-yl (isopropyl) or 2-methylprop-2-yl (t-butyl).In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene, optionally substituted C₂₋₁₂ alkenylene, optionallysubstituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂ cycloalkyleneor optionally substituted 3-12 membered heterocyclylene. In anotheraspect of this embodiment, L³ is optionally substituted C₃₋₁₂cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted C₄₋₆ cycloalkylene. In another aspect of this embodiment, L³is optionally substituted cyclohexylene. In another aspect of thisembodiment, L⁴ is —NH—. In another aspect of this embodiment, L⁴ isoptionally substituted C₁₋₁₂ alkylene. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y). In another aspect of this embodiment,X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachFr and R^(y) is independently methyl or ethyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹ isalkynylene, n is 0, and R³ is -(L³)_(p)-(L⁴)_(q)-(X³) wherein p is 1 andq is 2. In another aspect of this embodiment, L¹ is ethynylene. Inanother aspect of this embodiment, X¹ is optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl or optionally substituted C₃₋₁₂ cycloalkyl. In anotheraspect of this embodiment, X¹ is optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X¹ is optionally substituted C₃-C₇secondary or tertiary alkyl. In another aspect of this embodiment, X¹ isoptionally substituted C₃-C₅ cycloalkyl. In another aspect of thisembodiment, X¹ is prop-2-yl (isopropyl) or 2-methylprop-2-yl (t-butyl).In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene, optionally substituted C₂₋₁₂ alkenylene, optionallysubstituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂ cycloalkyleneor optionally substituted 3-12 membered heterocyclylene. In anotheraspect of this embodiment, L³ is optionally substituted C₃₋₁₂cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted C₄₋₆ cycloalkylene. In another aspect of this embodiment, L³is optionally substituted cyclohexylene. In another aspect of thisembodiment, L⁴ is —NH—. In another aspect of this embodiment, L⁴ isoptionally substituted C₁₋₁₂ alkylene. In another aspect of thisembodiment, one of L⁴ is —NH— and the other L⁴ is C₁₋₁₂ alkylene. Inanother aspect of this embodiment, -(L⁴)₂- is —NH—C₁₋₆ alkylene. Inanother aspect of this embodiment, X³ is —P(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —P(O)R^(x)R^(Y) wherein each R^(x) andR^(y) is independently ethyl or methyl. In another aspect of thisembodiment, X³ is —P(O)OR^(x)R^(y). In another aspect of thisembodiment, X³ is —P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R′ is independently methyl or ethyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹optionally substituted arylene, n is 0 and R³ is -(L³)_(p)-(L⁴)_(q)-(X³)wherein p is 1 and q is 0. In another aspect of this embodiment, L¹ isphenylene. In another aspect of this embodiment, X¹ is optionallysubstituted 3-14 membered heteroaryl. In another aspect of thisembodiment, X¹ is a 5-10 membered heteroaryl comprising one to threenitrogen atoms. In another aspect of this embodiment, L³ is optionallysubstituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂ alkenylene,optionally substituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene or optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene. In another aspect of this embodiment, L³ is optionallysubstituted ethylene. In another aspect of this embodiment, L³ isoptionally substituted C₃₋₁₂ cycloalkylene. In another aspect of thisembodiment, L³ is optionally substituted C₄₋₆ cycloalkylene. In anotheraspect of this embodiment, L³ is optionally substituted cyclohexylene.In another aspect of this embodiment, L³ is optionally substituted 3-12membered heterocyclylene. In another aspect of this embodiment, L³ isoptionally substituted 5-6 membered N-containing heterocyclene. Inanother aspect of this embodiment, L³ is piperidinylene. In anotheraspect of this embodiment, X³ is —P(O)R^(x)R^(y). In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x)(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In one embodiment of Formula I, R¹ is -(L¹)_(m)-(L²)_(n)-X¹, L¹optionally substituted arylene n is 1 and R³ is -(L³)_(p)-(L⁴)_(q)-(X³)wherein p is 1 and q is 0. In another aspect of this embodiment, L¹ isphenylene. In another aspect of this embodiment, L² is —NHC(O)—. Inanother aspect of this embodiment, X¹ is optionally substituted 3-14membered heteroaryl. In another aspect of this embodiment, X¹ isthiazolyl. In another aspect of this embodiment, L³ is optionallysubstituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂ alkenylene,optionally substituted C₂₋₁₂ alkynylene, optionally substituted C₃₋₁₂cycloalkylene or optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted C₁₋₁₂alkylene. In another aspect of this embodiment, L³ is optionallysubstituted ethylene. In another aspect of this embodiment, L³ isoptionally substituted C₃₋₁₂ cycloalkylene. In another aspect of thisembodiment, L³ is optionally substituted C₄₋₆ cycloalkylene. In anotheraspect of this embodiment, L³ is optionally substituted cyclohexylene.In another aspect of this embodiment, L³ is optionally substituted 3-12membered heterocyclylene. In another aspect of this embodiment, L³ isoptionally substituted 5-6 membered N-containing heterocyclene. Inanother aspect of this embodiment, L³ is piperidinylene. In anotheraspect of this embodiment, X³ is —P(O)R^(x)R^(y). In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently optionally substituted C₁₋₁₂ alkyl. In another aspect ofthis embodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In another aspect of this embodiment,R² is optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, or optionally substituted C₆₋₁₈ arylalkyl. In anotheraspect of this embodiment, R² is optionally substituted C₃₋₁₂cycloalkyl. In another aspect of this embodiment, R² is optionallysubstituted methylcyclohexyl. In another aspect of this embodiment, R²is optionally substituted methylcyclohexenyl. In another aspect of thisembodiment, R² is optionally substituted 4-methylcyclohexyl. In apreferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment, compounds of Formula I are represented bycompounds of Formula II:

or pharmaceutically acceptable salts and esters thereof, wherein:

R² is optionally substituted 4-methylcyclohexyl or optionallysubstituted methylcyclohexenyl;

X³ is selected from the group consisting of:

-   -   a) —P(O)R^(x)R^(y),    -   b)—P(O)R^(x)R^(y),    -   c)—OP(O)R^(x)R^(y),    -   d) —P(O)(OR^(x))(OR^(y));    -   e)

-   -   where each A is a 4 to 6 membered ring, including the depicted P        atom; and    -   f)

andthe remaining variables are defined as for Formula I.

in one embodiment of Formula II, R² is:

In one embodiment of Formula II, R² is:

In another embodiment of Formula II, R² is

In another embodiment of Formula II, R² is

In another embodiment of Formula II, p and q are each 0. In anotheraspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In a preferred aspect of thisembodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula II, p is 1 and q is 0. In anotheraspect of this embodiment, L³ is optionally substituted C₁₋₁₂ alkylene,optionally substituted C₂₋₁₂ alkenylene, optionally substituted C₂₋₁₂alkynylene, optionally substituted C₃₋₁₂ cycloalkylene or optionallysubstituted 3-12 membered heterocyclylene. In another aspect of thisembodiment, L³ is optionally substituted C₁₋₁₂ alkylene. In anotheraspect of this embodiment, L³ is optionally substituted ethylene. Inanother aspect of this embodiment, L³ is optionally substituted C₃₋₁₂cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted C₄₋₆ cycloalkylene. In another aspect of this embodiment, L³is optionally substituted cyclohexylene. In another aspect of thisembodiment, L³ is optionally substituted 3-12 membered heterocyclylene.In another aspect of this embodiment, L³ is optionally substituted 5-6membered N-containing heterocyclene. In another aspect of thisembodiment, L³ is piperidinylene. In another aspect of this embodiment,X³ is —P(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently ethyl ormethyl. In another aspect of this embodiment, X³ is —P(O)OR^(x)R^(y). Inanother aspect of this embodiment, X³ is —P(O)OR^(x)R^(y) wherein eachR^(x) and R^(y) is independently optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃). In anotheraspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is Hor C₁₋₆ alkyl. In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —OP(O)(CH₃)₂. In another aspect of this embodiment, X³is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(Y) is independentlymethyl, ethyl, or optionally substituted phenyl. In another aspect ofthis embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is —P(O)(OR^(x))(OR^(y)). In another aspect of this embodiment, X³ is—P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) is independentlymethyl or ethyl. In another aspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In a preferred aspect of thisembodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula II, p is 1 and q is 1. In anotheraspect of this embodiment, L³ is optionally substituted C₁₋₁₂ alkylene,optionally substituted C₂₋₁₂ alkenylene, optionally substituted C₂₋₁₂alkynylene, optionally substituted C₃₋₁₂ cycloalkylene or optionallysubstituted 3-12 membered heterocyclylene. In another aspect of thisembodiment, L³ is optionally substituted C₃₋₁₂ cycloalkylene. In anotheraspect of this embodiment, L³ is optionally substituted C₄₋₆cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted cyclohexylene. In another aspect of this embodiment, L⁴ is—NH—. In another aspect of this embodiment, L⁴ is optionally substitutedC₁₋₁₂ alkylene. In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independently optionallysubstituted C₁₋₁₂ alkyl. In another aspect of this embodiment, X³ is—P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is in dependently ethyl ormethyl. In another aspect of this embodiment, X³ is —P(O)OR^(x)R^(y). Inanother aspect of this embodiment, X³ is —P(O)OR^(x)R^(y) wherein eachR^(x) and R^(y) is independently optionally substituted C₁₋₁₂ alkyl. Inanother aspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃). In anotheraspect of this embodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is Hor C₁₋₆ alkyl. In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y). In another aspect of this embodiment, X³ is—OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —OP(O)(CH₃)₂. In another aspect of this embodiment, X³is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlymethyl, ethyl, or optionally substituted phenyl. In another aspect ofthis embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is —P(O)(OR^(x))(OR^(y)). In another aspect of this embodiment, X³ is—P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R′ is independently methylor ethyl. In another aspect of this embodiment, X³ is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In a preferred aspect of thisembodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment of Formula II, p is 1 and q is 2. In anotheraspect of this embodiment, L³ is optionally substituted C₁₋₁₂ alkylene,optionally substituted C₂₋₁₂ alkenylene, optionally substituted C₂₋₁₂alkynylene, optionally substituted C₃₋₁₂ cycloalkylene or optionallysubstituted 3-12 membered heterocyclylene. In another aspect of thisembodiment, L³ is optionally substituted C₃₋₁₂ cycloalkylene. In anotheraspect of this embodiment, L³ is optionally substituted C₄₋₆cycloalkylene. In another aspect of this embodiment, L³ is optionallysubstituted cyclohexylene. In another aspect of this embodiment, L⁴ is—NH—. In another aspect of this embodiment, L⁴ is optionally substitutedC₁₋₁₂ alkylene. In another aspect of this embodiment, one of L⁴ is —NH—and the other L⁴ is C₁₋₁₂ alkylene. In another aspect of thisembodiment, -(L⁴)₂- is —NH—C₁₋₆ alkylene. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y). In another aspect of this embodiment,X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently ethyl or methyl. In another aspect of this embodiment, X³is —P(O)OR^(x)R^(y). In another aspect of this embodiment, X³ is—P(O)OR^(x)R^(y) wherein each R^(x) and R^(y) is independentlyoptionally substituted C₁₋₁₂ alkyl. In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(CH₃) wherein R^(x) is H or C₁₋₆ alkyl.In another aspect of this embodiment, X³ is —OP(O)R^(x)R^(y). In anotheraspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) andR^(y) is independently optionally substituted C₁₋₁₂ alkyl. In anotheraspect of this embodiment, X³ is —OP(O)(CH₃)₂. In another aspect of thisembodiment, X³ is —OP(O)R^(x)R^(y) wherein each R^(x) and R^(y) isindependently methyl, ethyl, or optionally substituted phenyl. Inanother aspect of this embodiment, X³ is —OP(O)R^(x)R^(y) wherein eachR^(x) and R^(y) is independently methyl or ethyl. In another aspect ofthis embodiment, X³ is —P(O)(OR^(x))(OR^(y)). In another aspect of thisembodiment, X³ is —P(O)(OR^(x))(OR^(y)) wherein each R^(x) and R^(y) isindependently methyl or ethyl. In another aspect of this embodiment, X³is

wherein each A is a 4 to 6 membered ring, including the depicted P atom.In another aspect of this embodiment, X³ is

In another aspect of this embodiment, X³ is

wherein R^(y) is H or C₁₋₆ alkyl. In a preferred aspect of thisembodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another preferred aspect of this embodiment, R² is

In another embodiment, the compound of Formula I is selected from thegroup consisting of

or a pharmaceutically acceptable salt or ester thereof.

All documents referenced herein are each incorporated by reference intheir entirety for all purposes.

Definitions

Unless stated otherwise, the following terms and phrases as used hereinare intended to have the following meanings. The fact that a particularterm or phrase is not specifically defined should not be correlated toindefiniteness or lacking clarity, but rather terms herein are usedwithin their ordinary meaning. When trade names are used herein,applicants intend to independently include the tradename product and theactive pharmaceutical ingredient(s) of the tradename product.

The term “treating”, and grammatical equivalents thereof, when used inthe context of treating a disease, means slowing or stopping theprogression of a disease, or ameliorating at least one symptom of adisease, more preferably ameliorating more than one symptom of adisease. For example, treatment of a hepatitis C virus infection caninclude reducing the HCV viral load in an HCV infected human being,and/or reducing the severity of jaundice present in an HCV infectedhuman being.

“Alkyl” is hydrocarbon containing normal, secondary, tertiary or cycliccarbon atoms. For example, an alkyl group can have 1 to 20 carbon atoms(i.e., C₁-C₂₀ alkyl), 1 to 10 carbon atoms (i.e., C₁-C₁₀ alkyl), or 1 to6 carbon atoms (i.e., C₁-C₆ alkyl). Examples of suitable alkyl groupsinclude, but are not limited to, methyl (Me, —CH₃), ethyl (Et, —CH₂CH₃),1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr, i-propyl,—CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃), 2-methyl-1-propyl(i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl, —CH(CH₃)CH₂CH₃),2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl (n-pentyl,—CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, and octyl (—(CH₂)₇CH₃).

“Alkoxy” means a group having the formula —O-alkyl, in which an alkylgroup, as defined above, is attached to the parent molecule via anoxygen atom. The alkyl portion of an alkoxy group can have 1 to 20carbon atoms (i.e., C₁-C₂₀ alkoxy), 1 to 12 carbon atoms (i.e., C₁-C₁₂alkoxy), or 1 to 6 carbon atoms (i.e., C₁-C₆ alkoxy). Examples ofsuitable alkoxy groups include, but are not limited to, methoxy (—O—CH₃or —OMe), ethoxy (—OCH₂CH₃ or —OEt), t-butoxy (—O—C(CH₃)₃ or —OtBu), andthe like.

“Haloalkyl” is an alkyl group, as defined above, in which one or morehydrogen atoms of the alkyl group is replaced with a halogen atom. Thealkyl portion of a haloalkyl group can have 1 to 20 carbon atoms (i.e.,C₁—O₂₀ haloalkyl), 1 to 12 carbon atoms (i.e., C₁-C₁₂ haloalkyl), or 1to 6 carbon atoms (i.e., C₁-C₆ alkyl). Examples of suitable haloalkylgroups include, but are not limited to, —CF₃, —CHF₂, —CFH₂, —CH₂CF₃, andthe like.

“Alkenyl” is a hydrocarbon containing normal, secondary, tertiary, orcyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp2 double bond. For example, an alkenyl group can have 2to 20 carbon atoms (i.e., C₂-C₂₀ alkenyl), 2 to 12 carbon atoms (i.e.,C₂-C₁₂ alkenyl), or 2 to 6 carbon atoms (i.e., C₂-C₆ alkenyl). Examplesof suitable alkenyl groups include, but are not limited to, vinyl(—CH═CH₂), allyl (—CH₂CH═CH₂), cyclopentenyl (—C₅H₇), and 5-hexenyl(—CH₂CH₂CH₂CH₂CH═CH₂).

“Alkynyl” is a hydrocarbon containing normal, secondary, tertiary orcyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp triple bond. For example, an alkynyl group can have 2to 20 carbon atoms (i.e., C₂-C₂₀ alkynyl), 2 to 12 carbon atoms (i.e.,C₂-C₁₂ alkyne,), or 2 to 6 carbon atoms (i.e., C₂-C₆ alkynyl). Examplesof suitable alkynyl groups include, but are not limited to, acetylenic(—C≡CH), propargyl (—CH₂C≡CH), and the like.

“Alkylene” refers to a saturated, branched or straight chain radical oror cyclic hydrocarbon radical having two monovalent radical centersderived by the removal of two hydrogen atoms from the same or twodifferent carbon atoms of a parent alkane. For example, an alkylenegroup can have 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6carbon atoms. Typical alkylene radicals include, but are not limited to,methylene (—CH₂—), 1,1-ethylene (—CH(CH₃)—), 1,2-ethylene (—CH₂CH₂—),1,1-propylene (—CH(CH₂CH₃)—), 1,2-propylene (—CH₂CH(CH₃)—),1,3-propylene (—CH₂CH₂CH₂—), 1,4-butylene (—CH₂CH₂CH₂CH₂—), and thelike.

“Alkenylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical having two monovalent radical centers derivedby the removal of two hydrogen atoms from the same or two differentcarbon atoms of a parent alkene. For example, and alkenylene group canhave 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms.Typical alkenylene radicals include, but are not limited to,1,2-ethylene (—CH═CH—).

“Alkynylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical having two monovalent radical centers derivedby the removal of two hydrogen atoms from the same or two differentcarbon atoms of a parent alkyne. For example, an alkynylene group canhave 1 to 20 carbon atoms, 1 to 10 carbon atoms, or 1 to 6 carbon atoms.Typical alkynylene radicals include, but are not limited to, acetylenepropargyl (—C≡C—), propargyl (—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡C—).

“Aryl” means a monovalent aromatic hydrocarbon radical derived by theremoval of one hydrogen atom from a single carbon atom of a parentaromatic ring system. For example, an aryl group can have 6 to 20 carbonatoms, 6 to 14 carbon atoms, or 6 to 12 carbon atoms. Typical arylgroups include, but are not limited to, radicals derived from benzene(e.g., phenyl), substituted benzene, naphthalene, anthracene, biphenyl,and the like.

“Arylene” refers to an aryl as defined above having two monovalentradical centers derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent aryl. Typical aryleneradicals include, but are not limited to, phenylene.

“Arylalkyl” refers to an acyclic alkyl radical in which one of thehydrogen atoms bonded to a carbon atom, typically a terminal or sp3carbon atom, is replaced with an aryl radical. Typical arylalkyl groupsinclude, but are not limited to, benzyl, 2-phenylethan-1-yl,naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. The arylalkyl group can comprise6 to 20 carbon atoms, e.g., the alkyl moiety is 1 to 6 carbon atoms andthe aryl moiety is 6 to 14 carbon atoms.

“Cycloalkyl” refers to a saturated or partially unsaturated ring having3 to 7 carbon atoms as a monocycle, 7 to 12 carbon atoms as a bicycle,and up to about 20 carbon atoms as a polycycle. Monocyclic cycloalkylgroups have 3 to 6 ring atoms, still more typically 5 or 6 ring atoms.Bicyclic cycloalkyl groups have 7 to 12 ring atoms, e.g., arranged as abicycle (4,5), (5,5), (5,6) or (6,6) system, or 9 or 10 ring atomsarranged as a bicycle (5,6) or (6,6) system. Cycloalkyl groups includehydrocarbon mono-, bi-, and poly-cyclic rings, whether fused, bridged,or Spiro. Non-limiting examples of monocyclic cycloalkyls includecyclopropyl, cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl,1-cyclopent-2-enyl, 1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl,1-cyclohex-2-enyl and 1-cyclohex-3-enyl. Non-limiting examples ofbicyclo cycloalkyls includes naphthyl, tetrahydronapthalene, decalineand bicyclo[3.1.0]hex-6-yl and the like.

“Cycloalkylene” refers to a cycloalkyl as defined above having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent cycloalkyl.Typical cycloalkylene radicals include, but are not limited to,cyclopropylene, cyclobutylene, cyclopentylene and cyclohexylene.

“Halogen” refers to F, Cl, Br, or I.

As used herein the term “haloalkyl” refers to an alkyl group, as definedherein, that is substituted with at least one halogen. Examples ofbranched or straight chained “haloalkyl” groups as used herein include,but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, andt-butyl substituted independently with one or more halogens, forexample, fluoro, chloro, bromo, and iodo. The term “haloalkyl” should beinterpreted to include such substituents as perfluoroalkyl groups suchas —CF₃.

As used herein, the term “haloalkoxy” refers to a group —OR^(a), whereR^(a) is a haloalkyl group as herein defined. As non-limiting examples,haloalkoxy groups include —O(CH₂)F, —O(CH)F₂, and —OCF₃.

“Heterocycle” or “heterocyclyl” refers to a saturated or partiallysaturated cyclic group having from 1 to 14 carbon atoms and from 1 to 6heteroatoms selected from N, S, P, or O, and includes single ring andmultiple ring systems including, fused, bridged, and Spiro ring systems.“Heterocycle” or “heterocyclyl” as used herein includes by way ofexample and not limitation those heterocycles described in Paquette, LeoA.; Principles of Modern Heterocyclic Chemistry (W. A. Benjamin, NewYork, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; The Chemistryof Heterocyclic Compounds, A Series of Monographs” (John Wiley & Sons,New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and28; and J. Am. Chem. Soc. (1960) 82:5566. In one embodiment, the carbon,nitrogen, phosphorus, or sulfur atom(s) of the heterocyclic group may beoxidized to provide for C(═O), N-oxide, phosphinane oxide, sulfinyl, orsulfonyl moieties.

As one example, substituted heterocyclyls include, for example,heterocyclic rings substituted with any of the substituents disclosedherein including oxo groups. A non-limiting example of a carbonylsubstituted heterocyclyl is:

Examples of heterocycles include by way of example and not limitationdihydroypyridyl, tetrahydropyridyl (piperidyl), tetrahydrothiophenyl,sulfur oxidized tetrahydrothiophenyl, piperidinyl, 4-piperidonyl,pyrrolidinyl, azetidinyl, 2-pyrrolidonyl, tetrahydrofuranyl,decahydroquinolinyl, octahydroisoquinolinyl, pyranyl, morpholinyl, andbis-tetrahydrofuranyl:

“Heterocyclene” or “heterocyclylene” refers to a “heterocycle” or“heterocyclyl” as defined above having two monovalent radical centersderived by the removal of two hydrogen atoms from the same or twodifferent carbon atoms of a parent heterocycle, the removal of twohydrogen atoms from two nitrogen atoms of a parent heterocycle, or theremoval of a hydrogen atom from a nitrogen and the removal of a hydrogenatom from a carbon atome of a parent heterocycle. Non-limiting examplesof heterocyclene or heterocyclylenes are:

“Heteroaryl” refers to a monovalent aromatic heterocyclyl having atleast one heteroatom in the ring. Thus, “heteroaryl” refers to anaromatic group of from 1 to 14 carbon atoms and 1 to 6 heteroatomsselected from oxygen, nitrogen, sulfur, or phosphorus. For multiple ringsystems, by way of example, the term “heteroaryl” includes fused,bridged, and spiro ring systems having aromatic and non-aromatic rings.In one embodiment, the carbon, nitrogen, or sulfur ring atom(s) of theheteroaryl group may be oxidized to provide for C(═O), N-oxide,sulfinyl, or sulfonyl moieties.

Examples of heteroaryls include by way of example and not limitationpyridyl, thiazolyl, pyrimidinyl, furanyl, thienyl, pyrrolyl, pyrazolyl,imidazolyl, tetrazolyl, benzofuranyl, thianaphthalenyl, indolyl,quinolinyl, isoquinolinyl, benzimidazolyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl,thienyl, thianthrenyl, isobenzofuranyl, chromenyl, xanthenyl,phenoxathinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl,pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, 1H-indazoly, purinyl,4H-quinolizinyl, naphthyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl,pteridinyl, 4aH-carbazolyl, carbazolyl, β-carbolinyl, phenanthridinyl,acridinyl, pyrimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,furazanyl, phenoxazinyl, isochromanyl, chromanyl, imidazolidinyl,imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, indolinyl,isoindolinyl, quinuclidinyl, morpholinyl, oxazolidinyl, benzotriazolyl,benzisoxazolyl, oxindolyl, benzoxazolinyl, and isatinoyl.“Heterocyclylene” refers to a heterocyclyl, as defined herein, derivedby replacing a hydrogen atom from a carbon atom or heteroatom of aheterocyclyl, with an open valence. Similarly, “heteroarylene” refers toan aromatic heterocyclylene.

“Heterocyclylalkyl” refers to an acyclic alkyl radical in which one ofthe hydrogen atoms bonded to a carbon atom, typically a terminal or sp3carbon atom, is replaced with a heterocyclyl radical (i.e., aheterocyclyl-alkylene-moiety). Typical heterocyclylalkyl groups include,but are not limited to heterocyclyl-CH₂—, 2-(heterocyclyl)ethan-1-yl,and the like, wherein the “heterocyclyl” portion includes any of theheterocyclyl groups described above, including those described inPrinciples of Modern Heterocyclic Chemistry. One skilled in the art willalso understand that the heterocyclyl group can be attached to the alkylportion of the heterocyclyl alkyl by means of a carbon-carbon bond or acarbon-heteroatom bond, with the proviso that the resulting group ischemically stable. The heterocyclylalkyl group comprises 2 to 20 carbonatoms and 1-6 heteroatoms, e.g., the alkyl portion of theheterocyclylalkyl group comprises 1 to 6 carbon atoms and theheterocyclyl moiety comprises 1 to 14 carbon atoms. Examples ofheterocyclylalkyls include by way of example and not limitation5-membered sulfur, oxygen, phosphorus, and/or nitrogen containingheterocycles such as pyrrolidiylmethyl, 2-tetrahydrofuranylylethan-1-yl,and the like, 6-membered sulfur, oxygen, and/or nitrogen containingheterocycles such as piperidinylmethyl, morpholinylmethyl,piperidinylethyl, teterahydropyranylethyl, and the like.

“Heteroarylalkyl” refers to an alkyl group, as defined herein, in whicha hydrogen atom has been replaced with a heteroaryl group as definedherein. Non-limiting examples of heteroaryl alkyl include—CH₂-pyridinyl, —CH₂-pyrrolyl, —CH₂—oxazolyl, —CH₂-indolyl,—CH₂-isoindolyl, —CH₂-purinyl, —CH₂-furanyl, —CH₂-thienyl,—CH₂-benzofuranyl, —CH₂-benzothiophenyl, —CH₂-carbazolyl,—CH₂-imidazolyl, —CH₂-thiazolyl, —CH₂-isoxazolyl, —CH₂-pyrazolyl,—CH₂-isothiazolyl, —CH₂-quinolyl, —CH₂-isoquinolyl, —CH₂-pyridazyl,—CH₂-pyrimidyl, —CH₂-pyrazyl, —CH(CH₃)-pyridinyl, —CH(CH₃)-pyrrolyl,—CH(CH₃)-oxazolyl,

—CH(CH₃)-indolyl, —CH(CH₃)-isoindolyl, —CH(CH₃)-purinyl,—CH(CH₃)-furanyl, —CH(CH₃)-thienyl, —CH(CH₃)-benzofuranyl,—CH(CH₃)-benzothiophenyl, —CH(CH₃)— carbazolyl,

—CH(CH₃)-imidazolyl, —CH(CH₃)-thiazolyl, —CH(CH₃)-isoxazolyl,—CH(CH₃)-pyrazolyl, —CH(CH₃)-isothiazolyl, —CH(CH₃)-quinolyl,—CH(CH₃)-isoquinolyl, —CH(CH₃)-pyridazyl, —CH(CH₃)-pyrimidyl,—CH(CH₃)-pyrazyl, and the like.

The term “heterocyclyloxy” represents a heterocyclyl group attached tothe adjacent atom by an oxygen.

When there is a sulfur atom present, the sulfur atom can be at differentoxidation levels, namely, S, SO, SO₂, or SO₃. All such oxidation levelsare within the scope of the present invention.

When there is a phosphorus atom present, the phosphorus atom can be atdifferent oxidation levels, namely, POR^(a)R^(b)R^(c), PO₂R^(a)R^(b), orPO₃R^(a)R^(b), where Fr, R^(b), and R^(c) each independently is chosenfrom H, C₁₋₁₂ alkyl, C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₆₋₁₄ aryl, 3-12membered heterocycle, 3-18 membered heteroarylalkyl, C₆₋₁₈ arylalkyl; ortwo taken together (with or without oxygens) form a 5 to 10 memberedheterocycle. All such oxidation levels are within the scope of thepresent invention

The term “optionally substituted” in reference to a particular moiety ofthe compound of Formula I-II (e.g., an optionally substituted arylgroup) refers to a moiety wherein all substiutents are hydrogen orwherein one or more of the hydrogens of the moiety may be replaced bysubstituents such as those listed under the definition of “substituted”or as otherwise specified.

The term “substituted” in reference to alkyl, alkylene, aryl, arylalkyl,alkoxy, heterocyclyl, heteroaryl, carbocyclyl, etc., for example,“substituted alkyl”, “substituted alkylene”, “substituted aryl”,“substituted arylalkyl”, “substituted heterocyclyl”, and “substitutedcarbocyclyl” means alkyl, alkylene, aryl, arylalkyl, heterocyclyl,carbocyclyl respectively, in which one or more hydrogen atoms are eachindependently replaced with a non-hydrogen substituent. Divalent groupsmay also be similarly substituted. Unless otherwise indicated, typicalsubstituents include, but are not limited to, —X, —R^(b), —O, ═O,—OR^(b), —SR^(b), —S⁻, —NR^(b) ₂, —N⁺R^(b) ₃, ═NR^(b), —CX₃, —CN, —OCN,—SCN, —N═C═O, —NCS, —NO, —NO₂, ═N₂, —N₃, —NHC(═O)R^(b), —OC(═O)R^(b),—NHC(═O)NR^(b) ₂, —S(≡O)₂—, —S(═O)₂OH, —S(═O)₂R^(b), —OS(═O)₂OR^(b),—S(═O)₂NR^(b) ₂, —S(═O)R^(b), —OP(═O)(OR^(b))₂, —P(═O)(OR^(b))₂,—P(═O)(O⁻)₂, —P(═O)(OH)₂, —P(O)(OR^(b))(O⁻), —C(═O)R^(b), —C(═O)X,—C(S)R^(b), —C(O)OR^(b), —C(O)O⁻, —C(S)OR^(b), —C(O)SR^(b), —C(S)SR^(b),—C(O)NR^(b) ₂, —C(S)NR^(b) ₂, —C(═NR^(b))NR^(b) ₂, where each X isindependently a halogen: F, Cl, Br, or I; and each R^(b) isindependently H, alkyl, aryl, arylalkyl, a heterocycle, or a protectinggroup or prodrug moiety. Alkylene, alkenylene, and alkynylene groups mayalso be similarly substituted. Unless otherwise indicated, when the term“substituted” is used in conjunction with groups such as arylalkyl,which have two or more moieties capable of substitution, thesubstituents can be attached to the aryl moiety, the alkyl moiety, orboth.

Those skilled in the art will recognize that when moieties such as“alkyl”, “aryl”, “heterocyclyl”, etc. are substituted with one or moresubstituents, they could alternatively be referred to as “alkylene”,“arylene”, “heterocyclylene”, etc. moieties (i.e., indicating that atleast one of the hydrogen atoms of the parent “alkyl”, “aryl”,“heterocyclyl” moieties has been replaced with the indicatedsubstituent(s)). When moieties such as “alkyl”, “aryl”, “heterocyclyl”,etc. are referred to herein as “substituted” or are showndiagrammatically to be substituted (or optionally substituted, e.g.,when the number of substituents ranges from zero to a positive integer),then the terms “alkyl”, “aryl”, “heterocyclyl”, etc. are understood tobe interchangeable with “alkylene”, “arylene”, “heterocyclylene”, etc.

As will be appreciated by those skilled in the art, the compounds of thepresent invention may exist in solvated or hydrated form. The scope ofthe present invention includes such forms. Again, as will be appreciatedby those skilled in the art, the compounds may be capable ofesterification. The scope of the present invention includes esters andother physiologically functional derivatives. The scope of the presentinvention includes prodrug forms of the compound herein described.

“Ester” means any ester of a compound in which any of the —COOHfunctions of the molecule is replaced by a —C(O)OR function, or in whichany of the —OH functions of the molecule are replaced with a —OC(O)Rfunction, in which the R moiety of the ester is any carbon-containinggroup which forms a stable ester moiety, including but not limited toalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl,heterocyclyl, heterocyclylalkyl and substituted derivatives thereof.

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates the drug substance, i.e.,active ingredient, as a result of spontaneous chemical reaction(s),enzyme catalyzed chemical reaction(s), photolysis, and/or metabolicchemical reaction(s). A prodrug is thus a covalently modified analog orlatent form of a therapeutically active compound. Non-limiting examplesof prodrugs include ester moieties, quaternary ammonium moieties, glycolmoieties, and the like.

One skilled in the art will recognize that substituents and othermoieties of the compounds of Formula I or II should be selected in orderto provide a compound which is sufficiently stable to provide apharmaceutically useful compound which can be formulated into anacceptably stable pharmaceutical composition. Compounds of Formula I orII which have such stability are contemplated as falling within thescope of the present invention.

As will be appreciated by those skilled in the art, the compounds of thepresent invention may contain one or more chiral centers. The scope ofthe present invention includes such forms. Again, as will be appreciatedby those skilled in the art, the compound is capable of esterification.The scope of the present invention includes esters and otherphysiologically functional derivatives. In addition, the scope of thepresent invention includes prodrug forms of the compound hereindescribed.

A compound of Formula I-II and its pharmaceutically acceptable salts mayexist as different polymorphs or pseudopolymorphs. As used herein,crystalline polymorphism means the ability of a crystalline compound toexist in different crystal structures. Polymorphism generally can occuras a response to changes in temperature, pressure, or both. Polymorphismcan also result from variations in the crystallization process.Polymorphs can be distinguished by various physical characteristicsknown in the art such as x-ray diffraction patterns, solubility, andmelting point. The crystalline polymorphism may result from differencesin crystal packing (packing polymorphism) or differences in packingbetween different conformers of the same molecule (conformationalpolymorphism). As used herein, crystalline pseudopolymorphism means theability of a hydrate or solvate of a compound to exist in differentcrystal structures. The pseudopolymorphs of the instant invention mayexist due to differences in crystal packing (packing pseudopolymorphism)or due to differences in packing between different conformers of thesame molecule (conformational pseudopolymorphism). The instant inventioncomprises all polymorphs and pseudopolymorphs of the compounds ofFormula I-II and their pharmaceutically acceptable salts.

A compound of Formula I-II and its pharmaceutically acceptable salts mayalso exist as an amorphous solid. As used herein, an amorphous solid isa solid in which there is no long-range order of the positions of theatoms in the solid. This definition applies as well when the crystalsize is two nanometers or less. Additives, including solvents, may beused to create the amorphous forms of the instant invention. The instantinvention comprises all amorphous forms of the compounds of Formula I-IIand their pharmaceutically acceptable salts.

Certain of the compounds described herein contain one or more chiralcenters, or may otherwise be capable of existing as multiplestereoisomers. The scope of the present invention includes mixtures ofstereoisomers as well as purified enantiomers orenantiomerically/diastereomerically enriched mixtures. Also includedwithin the scope of the invention are the individual isomers of thecompounds represented by the formulae of the present invention, as wellas any wholly or partially equilibrated mixtures thereof. The presentinvention also includes the individual isomers of the compoundsrepresented by the formulas above as mixtures with isomers thereof inwhich one or more chiral centers are inverted.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g., melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as electrophoresis and chromatography.

“Enantiomers” refer to stereoisomers of a compound which arenon-superimposable mirror images of one another.

“Atropisomers” refer to stereoisomers of a compound resulting fromhindered rotation about single bonds where the steric strain barrier torotation is high enough to allow for the isolation of the individualconformer. Atropisomers display axial chirality. Atropisomers may beequilibrated thermally and the interconversion barrier may be measuredkinetically. Atropisomerism may occur apart from the presence of otherforms of chiral isomerism. Thus, as illustrated, the depicted nitrogenatom is planar and compound of Formula I is capable of existing asatropisomers:

In one embodiment of the present invention, the compounds exist in aconformeric form of Formula Ia:

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork.

Many organic compounds exist in optically active forms, i.e., they havethe ability to rotate the plane of plane-polarized light. In describingan optically active compound, the prefixes D and L or R and S are usedto denote the absolute configuration of the molecule about its chiralcenter(s). The prefixes d and I or (+) and (−) are employed to designatethe sign of rotation of plane-polarized light by the compound, with (−)or 1 meaning that the compound is levorotatory. A compound prefixed with(+) or d is dextrorotatory.

A specific stereoisomer may also be referred to as an enantiomer, and amixture of such isomers is often called an enantiomeric mixture. A 50:50mixture of enantiomers is referred to as a racemic mixture or aracemate, which may occur where there has been no stereoselection orstereospecificity in a chemical reaction or process. The terms “racemicmixture” and “racemate” refer to an equimolar mixture of twoenantiomeric species, devoid of optical activity.

The present invention includes a salt or solvate of the compounds hereindescribed, including combinations thereof such as a solvate of a salt.The compounds of the present invention may exist in solvated, forexample hydrated, as well as unsolvated forms, and the present inventionencompasses all such forms.

Typically, but not absolutely, the salts of the present invention arepharmaceutically acceptable salts. Salts encompassed within the term“pharmaceutically acceptable salts” refer to non-toxic salts of thecompounds of this invention.

Examples of suitable pharmaceutically acceptable salts include inorganicacid addition salts such as chloride, bromide, sulfate, phosphate, andnitrate; organic acid addition salts such as acetate, galactarate,propionate, succinate, lactate, glycolate, malate, tartrate, citrate,maleate, fumarate, methanesulfonate, p-toluenesulfonate, and ascorbate;salts with acidic amino acid such as aspartate and glutamate; alkalimetal salts such as sodium salt and potassium salt; alkaline earth metalsalts such as magnesium salt and calcium salt; ammonium salt; organicbasic salts such as trimethylamine salt, triethylamine salt, pyridinesalt, picoline salt, dicyclohexylamine salt, andN,N′-dibenzylethylenediamine salt; and salts with basic amino acid suchas lysine salt and arginine salt. The salts may be in some caseshydrates or ethanol solvates.

The modifier “about” used in connection with a quantity is inclusive ofthe stated value and has the meaning dictated by the context (e.g.,includes the degree of error associated with measurement of theparticular quantity).

Whenever a compound described herein is substituted with more than oneof the same designated group, e.g., “R” or “R¹”, then it will beunderstood that the groups may be the same or different, i.e., eachgroup is independently selected. Wavy lines,

indicate the site of covalent bond attachments to the adjoiningsubstructures, groups, moieties, or atoms.

The compounds of the invention can also exist as tautomeric isomers incertain cases. Although only one delocalized resonance structure may bedepicted, all such forms are contemplated within the scope of theinvention. For example, ene-amine tautomers can exist for purine,pyrimidine, imidazole, guanidine, amidine, and tetrazole systems and alltheir possible tautomeric forms are within the scope of the invention.

Selected substituents comprising the compounds of Formula I-II may bepresent to a recursive degree. In this context, “recursive substituent”means that a substituent may recite another instance of itself. Themultiple recitations may be direct or indirect through a sequence ofother substituents. Because of the recursive nature of suchsubstituents, theoretically, a large number of compounds may be presentin any given embodiment. One of ordinary skill in the art of medicinalchemistry understands that the total number of such substituents isreasonably limited by the desired properties of the compound intended.Such properties include, by way of example and not limitation, physicalproperties such as molecular weight, solubility or log P, applicationproperties such as activity against the intended target, and practicalproperties such as ease of synthesis. Recursive substituents may be anintended aspect of the invention. One of ordinary skill in the art ofmedicinal chemistry understands the versatility of such substituents. Tothe degree that recursive substituents are present in an embodiment ofthe invention, they may recite another instance of themselves, 0, 1, 2,3, or 4 times.

The compounds of Formula I-II also include molecules that incorporateisotopes of the atoms specified in the particular molecules.Non-limiting examples of these isotopes include D, T, ¹⁴C, ¹³C and ¹⁵N.

Protecting Groups

In the context of the present invention, protecting groups includeprodrug moieties and chemical protecting groups.

Protecting groups are available, commonly known and used, and areoptionally used to prevent side reactions with the protected groupduring synthetic procedures, i.e. routes or methods to prepare thecompounds of the invention. For the most part the decision as to whichgroups to protect, when to do so, and the nature of the chemicalprotecting group “PG” will be dependent upon the chemistry of thereaction to be protected against (e.g., acidic, basic, oxidative,reductive or other conditions) and the intended direction of thesynthesis. The PG groups do not need to be, and generally are not, thesame if the compound is substituted with multiple PG. In general, PGwill be used to protect functional groups such as carboxyl, hydroxyl,thio, or amino groups and to thus prevent side reactions or to otherwisefacilitate the synthetic efficiency. The order of deprotection to yieldfree, deprotected groups is dependent upon the intended direction of thesynthesis and the reaction conditions to be encountered, and may occurin any order as determined by the artisan.

Various functional groups of the compounds of the invention may beprotected. For example, protecting groups for —OH groups (whetherhydroxyl, carboxylic acid, phosphonic acid, or other functions) include“ether- or ester-forming groups”. Ether- or ester-forming groups arecapable of functioning as chemical protecting groups in the syntheticschemes set forth herein. However, some hydroxyl and thio protectinggroups are neither ether- nor ester-forming groups, as will beunderstood by those skilled in the art, and are included with amides,discussed below.

A very large number of hydroxyl protecting groups and amide-forminggroups and corresponding chemical cleavage reactions are described inProtective Groups in Organic Synthesis, Theodora W. Greene and Peter G.M. Wuts (John Wiley & Sons, Inc., New York, 1999, ISBN 0-471-16019-9)(“Greene”). See also Kocienski, Philip J.; Protecting Groups (GeorgThieme Verlag Stuttgart, New York, 1994), which is incorporated byreference in its entirety herein. In particular Chapter 1, ProtectingGroups: An Overview, pages 1-20, Chapter 2, Hydroxyl Protecting Groups,pages 21-94, Chapter 3, Diol Protecting Groups, pages 95-117, Chapter 4,Carboxyl Protecting Groups, pages 118-154, Chapter 5, CarbonylProtecting Groups, pages 155-184. For protecting groups for carboxylicacid, phosphonic acid, phosphonate, sulfonic acid and other protectinggroups for acids see Greene as set forth below. Such groups include byway of example and not limitation, esters, amides, hydrazides, and thelike.

Ether- and Ester-Forming Protecting Groups

Ester-forming groups include: (1) phosphonate ester-forming groups, suchas phosphonamidate esters, phosphorothioate esters, phosphonate esters,and phosphon-bis-amidates; (2) carboxyl ester-forming groups, and (3)sulphur ester-forming groups, such as sulphonate, sulfate, andsulfinate.

Metabolites of the Compounds of the Invention

Also falling within the scope of this invention are the in vivometabolic products of the compounds described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,esterification and the like of the administered compound, primarily dueto enzymatic processes. Accordingly, the invention includes compoundsproduced by a process comprising contacting a compound of this inventionwith a mammal for a period of time sufficient to yield a metabolicproduct thereof. Such products typically are identified by preparing aradiolabelled C¹⁴ or H³) compound of the invention, administering itparenterally in a detectable dose (e.g., greater than about 0.5 mg/kg)to an animal such as rat, mouse, guinea pig, monkey, or to man, allowingsufficient time for metabolism to occur (typically about 30 seconds to30 hours) and isolating its conversion products from the urine, blood orother biological samples. These products are easily isolated since theyare labeled (others are isolated by the use of antibodies capable ofbinding epitopes surviving in the metabolite). The metabolite structuresare determined in conventional fashion, e.g., by MS or NMR analysis. Ingeneral, analysis of metabolites is done in the same way as conventionaldrug metabolism studies well-known to those skilled in the art. Theconversion products, so long as they are not otherwise found in vivo,are useful in diagnostic assays for therapeutic dosing of the compoundsof the invention even if they possess no anti-infective activity oftheir own.

The definitions and substituents for various genus and subgenus of thepresent compounds are described and illustrated herein. It should beunderstood by one skilled in the art that any combination of thedefinitions and substituents described above should not result in aninoperable species or compound. “Inoperable species or compounds” meanscompound structures that violates relevant scientific principles (suchas, for example, a carbon atom connecting to more than four covalentbonds) or compounds too unstable to permit isolation and formulationinto pharmaceutically acceptable dosage forms.

Pharmaceutical Formulations

The compounds of this invention are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic. All formulations will optionally contain excipientssuch as those set forth in the Handbook of Pharmaceutical Excipients(1986), herein incorporated by reference in its entirety. Excipientsinclude ascorbic acid and other antioxidants, chelating agents such asEDTA, carbohydrates such as dextrin, hydroxyalkylcellulose,hydroxyalkylmethylcellulose, stearic acid and the like. The pH of theformulations ranges from about 3 to about 11, but is ordinarily about 7to 10.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations of the invention, both for veterinary and for human use,comprise at least one active ingredient, together with one or moreacceptable carriers and optionally other therapeutic ingredients. Thecarrier(s) must be “acceptable” in the sense of being compatible withthe other ingredients of the formulation and physiologically innocuousto the recipient thereof.

The formulations include those suitable for the foregoing administrationroutes. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Techniques and formulations generally are found in Remington'sPharmaceutical Sciences (Mack Publishing Co., Easton, Pa.), hereinincorporated by reference in its entirety. Such methods include the stepof bringing into association the active ingredient with the carrierwhich constitutes one or more accessory ingredients. In general theformulations are prepared by uniformly and intimately bringing intoassociation the active ingredient with liquid carriers or finely dividedsolid carriers or both, and then, if necessary, shaping the product.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or a suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas a powder or granules, optionally mixed with a binder, lubricant,inert diluent, preservative, surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered active ingredient moistened with an inert liquid diluent. Thetablets may optionally be coated or scored and optionally are formulatedso as to provide slow or controlled release of the active ingredient.

For administration to the eye or other external tissues e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w (including active ingredient(s) in a range between 0.1%and 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.),preferably 0.2 to 15% w/w and most preferably 0.5 to 10% w/w. Whenformulated in an ointment, the active ingredients may be employed witheither a paraffinic or a water-miscible ointment base. Alternatively,the active ingredients may be formulated in a cream with an oil-in-watercream base.

If desired, the aqueous phase of the cream base may include, forexample, at least 30% w/w of a polyhydric alcohol, i.e. an alcoholhaving two or more hydroxyl groups such as propylene glycol, butane1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol(including PEG 400) and mixtures thereof. The topical formulations maydesirably include a compound which enhances absorption or penetration ofthe active ingredient through the skin or other affected areas. Examplesof such dermal penetration enhancers include dimethyl sulphoxide andrelated analogs.

The oily phase of the emulsions of this invention may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the invention include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties. The cream should preferablybe a non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as di-isoadipate,isocetyl stearate, propylene glycol diester of coconut fatty acids,isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters known asCrodamol CAP may be used, the last three being preferred esters. Thesemay be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils are used.

Pharmaceutical formulations according to the present invention compriseone or more compounds of the invention together with one or morepharmaceutically acceptable carriers or excipients and optionally othertherapeutic agents. Pharmaceutical formulations containing the activeingredient may be in any form suitable for the intended method ofadministration. When used for oral use for example, tablets, troches,lozenges, aqueous or oil suspensions, dispersible powders or granules,emulsions, hard or soft capsules, syrups or elixirs may be prepared.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsincluding sweetening agents, flavoring agents, coloring agents andpreserving agents, in order to provide a palatable preparation. Tabletscontaining the active ingredient in admixture with non-toxicpharmaceutically acceptable excipient which are suitable for manufactureof tablets are acceptable. These excipients may be, for example, inertdiluents, such as calcium or sodium carbonate, lactose, lactosemonohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as maize starch,or alginic acid; binding agents, such as cellulose, microcrystallinecellulose, starch, gelatin or acacia; and lubricating agents, such asmagnesium stearate, stearic acid or talc. Tablets may be uncoated or maybe coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions.

Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, hydroxypropyl methylcelluose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia,and dispersing or wetting agents such as a naturally occurringphosphatide (e.g., lecithin), a condensation product of an alkyleneoxide with a fatty acid (e.g., polyoxyethylene stearate), a condensationproduct of ethylene oxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth herein, andflavoring agents may be added to provide a palatable oral preparation.These compositions may be preserved by the addition of an antioxidantsuch as ascorbic acid.

Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachis oil, a mineral oil, such as liquid paraffin, ora mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion may also contain sweetening and flavoring agents. Syrups andelixirs may be formulated with sweetening agents, such as glycerol,sorbitol or sucrose. Such formulations may also contain a demulcent, apreservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned herein. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye dropswherein the active ingredient is dissolved or suspended in a suitablecarrier, especially an aqueous solvent for the active ingredient. Theactive ingredient is preferably present in such formulations in aconcentration of 0.5 to 20%, advantageously 0.5 to 10% particularlyabout 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 μm (includingparticle sizes in a range between 0.1 and 500 μm in increments such as0.5 μm, 1 μm, 30 μm, 35 μm, etc.), which is administered by rapidinhalation through the nasal passage or by inhalation through the mouthso as to reach the alveolar sacs. Suitable formulations include aqueousor oily solutions of the active ingredient. Formulations suitable foraerosol or dry powder administration may be prepared according toconventional methods and may be delivered with other therapeutic agentssuch as compounds heretofore used in the treatment or prophylaxis ofinfections as described herein.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

The formulations are presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this invention may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

Compounds of the invention can also be formulated to provide controlledrelease of the active ingredient to allow less frequent dosing or toimprove the pharmacokinetic or toxicity profile of the activeingredient. Accordingly, the invention also provided compositionscomprising one or more compounds of the invention formulated forsustained or controlled release.

The effective dose of an active ingredient depends at least on thenature of the condition being treated, toxicity, whether the compound isbeing used prophylactically (lower doses) or against an active viralinfection, the method of delivery, and the pharmaceutical formulation,and will be determined by the clinician using conventional doseescalation studies. The effective dose can be expected to be from about0.0001 to about 100 mg/kg body weight per day; typically, from about0.01 to about 10 mg/kg body weight per day; more typically, from about0.01 to about 5 mg/kg body weight per day; most typically, from about0.05 to about 0.5 mg/kg body weight per day. For example, the dailycandidate dose for an adult human of approximately 70 kg body weightwill range from 1 mg to 1000 mg, preferably between 5 mg and 500 mg, andmay take the form of single or multiple doses.

In yet another-embodiment, the present application disclosespharmaceutical compositions comprising a compound of Formula I or apharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier or exipient.

Routes of Administration

One or more compounds of the invention (herein referred to as the activeingredients) are administered by any route appropriate to the conditionto be treated. Suitable routes include oral, rectal, nasal, topical(including buccal and sublingual), vaginal and parenteral (includingsubcutaneous, intramuscular, intravenous, intradermal, intrathecal andepidural), and the like. It will be appreciated that the preferred routemay vary with for example the condition of the recipient. An advantageof the compounds of this invention is that they are orally bioavailableand can be dosed orally.

Combination Therapy, Including HCV Combination Therapy

In another embodiment, the compounds of the present invention may becombined with one or more active agent. Non-limiting examples ofsuitable combinations include combinations of one or more compounds ofthe present invention with one or more interferons, ribavirin or itsanalogs, HCV NS3 protease inhibitors, NS5a inhibitors, alpha-glucosidase1 inhibitors, hepatoprotectants, mevalonate decarboxylase antagonists,antagonists of the renin-angiotensin system, other anti-fibrotic agents,endothelin antagonists, nucleoside or nucleotide inhibitors of HCV NS5Bpolymerase, non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5Ainhibitors, TLR-7 agonists, cyclophillin inhibitors, HCV IRESinhibitors, pharmacokinetic enhancers and other drugs for treating HCV;or mixtures thereof.

More specifically, one or more compounds of the present invention may becombined with one or more compounds selected from the group consistingof

1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron), pegylatedrIFN-alpha 2a (Pegasys), rIFN-alpha 2b (Intron A), rIFN-alpha 2a(Roferon-A), interferon alpha (MOR-22, OPC-18, Alfaferone, Alfanative,Multiferon, subalin), interferon alfacon-1 (Infergen), interferonalpha-n1 (Wellferon), interferon alpha-n3 (Alferon), interferon-beta(Avonex, DL-8234), interferon-omega (omega DUROS, Biomed 510),albinterferon alpha-2b (Albuferon), IFN alpha XL, BLX-883 (Locteron),DA-3021, glycosylated interferon alpha-2b (AVI-005), PEG-Infergen,PEGylated interferon lambda (PEGylated IL-29), and belerofon,

2) ribavirin and its analogs, e.g., ribavirin (Rebetol, Copegus), andtaribavirin (Viramidine),

3) HCV NS3 protease inhibitors, e.g., boceprevir (SCH-503034 SCH-7),telaprevir (VX-950), VX-813, TMC-435 (TMC435350), ABT-450, BI-201335,BI-1230, MK-7009, SCH-900518, VBY-376, VX-500, GS-9256, GS-9451,BMS-790052, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530, YH5531, andITMN-191 (R-7227),

4) alpha-glucosidase 1 inhibitors, e.g., celgosivir (MX-3253), Miglitol,and UT-231B,

5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738, GS-9450(LB-84451), silibilin, and MitoQ,

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, e.g.,R1626, R7128 (R4048), IDX184, IDX-102, PSI-7851, BCX-4678,valopicitabine (NM-283), and MK-0608,

7) non-nucleoside inhibitors of HCV NS5B polymerase, e.g., filibuvir(PF-868554), ABT-333, ABT-072, BI-207127, VCH-759, VCH-916, JTK-652,MK-3281, VBY-708, VCH-222, A848837, ANA-598, GL60667, GL59728, A-63890,A-48773, A-48547, BC-2329, VCH-796 (nesbuvir), GSK625433, BILN-1941,XTL-2125, and GS-9190,

8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), AZD-7295 (A-689), andBMS-790052,

9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773, ANA-975,AZD-8848 (DSP-3025), PF-04878691, and SM-360320,

10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and NIM811,

11) HCV IRES inhibitors, e.g., MCI-067,

12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452, PF-4194477,TMC-41629, GS-9350, GS-9585, and roxythromycin,

13) other drugs for treating HCV, e.g., thymosin alpha 1 (Zadaxin),nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir, GI-5005,XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOV-205, tarvacin,EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032, BMS-791325,Bavituximab, MDX-1106 (ONO-4538), Oglufanide, FK-788, and VX-497(merimepodib)

14) mevalonate decarboxylase antagonists, e.g., statins, HMGCoA synthaseinhibitors (e.g., hymeglusin), squalene synthesis inhibitors (e.g.,zaragozic acid);

15) angiotensin II receptor antagonists, e.g., losartan, irbesartan,olmesartan, candesartan, valsartan, telmisartan, eprosartan;

16) angiotensin-converting enzyme inhibitors, e.g., captopril,zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril,benazepril, fosinopril;

17) other anti-fibrotic agents, e.g., amiloride and

18) endothelin antagonists, e.g. bosentan and ambrisentan.

In yet another embodiment, the present application disclosespharmaceutical compositions comprising a compound of the presentinvention, or a pharmaceutically acceptable salt thereof, in combinationwith at least one additional active agent, and a pharmaceuticallyacceptable carrier or excipient. In yet another embodiment, the presentapplication provides a combination pharmaceutical agent with two or moretherapeutic agents in a unitary dosage form. Thus, it is also possibleto combine any compound of the invention with one or more other activeagents in a unitary dosage form.

The combination therapy may be administered as a simultaneous orsequential regimen. When administered sequentially, the combination maybe administered in two or more administrations.

Co-administration of a compound of the invention with one or more otheractive agents generally refers to simultaneous or sequentialadministration of a compound of the invention and one or more otheractive agents, such that therapeutically effective amounts of thecompound of the invention and one or more other active agents are bothpresent in the body of the patient.

Co-administration includes administration of unit dosages of thecompounds of the invention before or after administration of unitdosages of one or more other active agents, for example, administrationof the compounds of the invention within seconds, minutes, or hours ofthe administration of one or more other active agents. For example, aunit dose of a compound of the invention can be administered first,followed within seconds or minutes by administration of a unit dose ofone or more other active agents. Alternatively, a unit dose of one ormore other active agents can be administered first, followed byadministration of a unit dose of a compound of the invention withinseconds or minutes. In some cases, it may be desirable to administer aunit dose of a compound of the invention first, followed, after a periodof hours (e.g., 1-12 hours), by administration of a unit dose of one ormore other active agents. In other cases, it may be desirable toadminister a unit dose of one or more other active agents first,followed, after a period of hours (e.g., 1-12 hours), by administrationof a unit dose of a compound of the invention.

The combination therapy may provide “synergy” and “synergistic effect”,i.e. the effect achieved when the active ingredients used together isgreater than the sum of the effects that results from using thecompounds separately. A synergistic effect may be attained when theactive ingredients are: (1) co-formulated and administered or deliveredsimultaneously in a combined formulation; (2) delivered by alternationor in parallel as separate formulations; or (3) by some other regimen.When delivered in alternation therapy, a synergistic effect may beattained when the compounds are administered or delivered sequentially,e.g., in separate tablets, pills or capsules, or by different injectionsin separate syringes. In general, during alternation therapy, aneffective dosage of each active ingredient is administered sequentially,i.e. serially, whereas in combination therapy, effective dosages of twoor more active ingredients are administered together.

As will be appreciated by those skilled in the art, when treating aviral infection such as HCV, such treatment may be characterized in avariety of ways and measured by a variety of endpoints. The scope of thepresent invention is intended to encompass all such characterizations.

SYNTHETIC EXAMPLES

Certain abbreviations and acronyms are used in describing theexperimental details. Although most of these would be understood by oneskilled in the art, Table 1 contains a list of many of theseabbreviations and acronyms.

TABLE 1 List of abbreviations and acronyms. Abbreviation Meaning Ac₂Oacetic anhydride AIBN 2,2′-azobis(2-methylpropionitrile) Bn benzyl BnBrbenzylbromide BSA bis(trimethylsilyl)acetamide BzCl benzoyl chloride CDIcarbonyl diimidazole DABCO 1,4-diazabicyclo[2.2.2]octane DBN1,5-diazabicyclo[4.3.0]non-5-ene DDQ2,3-dichloro-5,6-dicyano-1,4-benzoquinone DBU1,5-diazabicyclo[5.4.0]undec-5-ene DCA dichloroacetamide DCCdicyclohexylcarbodiimide DCM dichloromethane DMAP4-dimethylaminopyridine DME 1,2-dimethoxyethane DMTCl dimethoxytritylchloride DMSO dimethylsulfoxide DMTr 4,4′-dimethoxytrityl DMFdimethylformamide EtOAc ethyl acetate ESI electrospray ionization HMDShexamethyldisilazane HPLC High pressure liquid chromatography LDAlithium diisopropylamide LRMS low resolution mass spectrum MCPBAmeta-chloroperbenzoic acid MeCN acetonitrile MeOH methanol MMTC monomethoxytrityl chloride m/z or m/e mass to charge ratio MH⁺ mass plus 1MH⁻ mass minus 1 MsOH methanesulfonic acid MS or ms mass spectrum NBSN-bromosuccinimide Ph phenyl rt or r.t. room temperature TBAFtetrabutylammonium fluoride TMSCl chlorotrimethylsilane TMSBrbromotrimethylsilane TMSI iodotrimethylsilane TMSOTf(trimethylsilyl)trifluoromethylsulfonate TEA triethylamine TBAtributylamine TBAP tributylammonium pyrophosphate TBSClt-butyldimethylsilyl chloride TEAB triethylammonium bicarbonate TFAtrifluoroacetic acid TLC or tlc thin layer chromatography Trtriphenylmethyl Tol 4-methylbenzoyl Turbo Grignard 1:1 mixture ofisopropylmagnesium chloride and lithium chloride δ parts per milliondown field from tetramethylsilaneGeneral Schemes I

The compounds of this invention may be synthesized by routes with keybond-forming steps as indicated in Scheme A, below, in which thecarboxylate substituent “R” indicates either a protecting group such asan alkyl ester (where applicable), or the free acid itself. Alkyl esterprotecting groups are conveniently removed by saponification with analkali metal hydroxide in a protic solvent such as water or an alcohol,and may be facilitated by use of ethereal solvent mixtures and/orheating. Alternatively they may be removed by dealkylation throughheating with an alkali metal halide in an aprotic solvent. Similarmanipulations may be performed on phosphinate-contaning intermediates.As a non-limiting example, phosphinoyl esters are convenientlydealkylated by heating with a Lewis acid such as iodotrimethylsilane,optionally in the presence of a base such as 2,6-dimethyl lutidine andan inert aprotic solvent such as acetonitrile.

When X³ indicates a group that is attached to L³ or L⁴ via a phosphorusatom, R³ may conveniently be introduced via reductive amination using aphosphorus-containing carbonyl reagent together with an appropriate3-aminothiophene, and mediated by a reducing agent such as phenylsilanein the presence of a Lewis acid such as dibutyl tin dichloride in aninert ethereal solvent. Alternatively the intermediate secondary aminemay be generated by coupling of a phosphorus-containing primary aminewith a 3-halothiophene in a reaction that may be catalyzed by Pd (J.Org. Chem., 2000, 65, 1158-1174, herein incorporated by reference withregard to such synthesis). The amine is then converted to the amide byacylation with a carboxylic acid derivative such as an acyl chloride oranhydride in the presence of a base such as pyridine or a tertiary aminein an inert solvent such as dichloromethane.

In other cases it is convenient to perform synthetic manipulations onthe side chain R³ with the rest of the molecule fully assembled. Thus,for example, if L³ is to contain a secondary amine, reductive aminationwith a phosphorus-containing aldehyde or ketone in the presence of areducing agent, such as sodium triacetoxyborohydride or sodiumcyanoborohydride, in a dipolar aprotic solvent generates the desiredtarget molecule directly. Alternatively, if the phosphorus atom isconnected to L³ or L⁴ via an oxygen atom (as when X³ is group c)), therequisite alcohol may be treated with a phosphinoyl chloride reagent inthe presence of a tertiary amine base. On the other hand, if thephosphorus atom is connected via a carbon atom bearing a hydroxyl group,the high nucleophilicity of phosphinates may be exploited to generatethe desired C—P bond through addition to a carbonyl group.

Experimentals

Example 1 Compound 1:5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-(dimethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

A mixture of5-(3,3-dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (100 mg, 0.224 mmol) and triethylamine (278 μL, 2.02 mmol) in DCM(1 mL) at 0° C. was treated with dimethylphosphinic chloride (150 mg,1.34 mmol) in two portions. The mixture was warm to room temperature andstirred for several hours.

Ethyl acetate and water was added and the aqueous layer was carefullyquenched with citric acid (10% aqueous solution, 2-3 mL). The mixturewas extracted with ethyl acetate (2×30 mL). The combined organic layerswere dried over sodium sulfate, filtered and concentrated. The residuewas purified by RP HPLC using a C18 column with a gradient of 0.1%TFA-H₂O, 0.1% TFA-acetonitrile, to provide 70 mg (60%) of desiredproduct. MS (m/z) 522.2 [M+H]⁺; HPLC retention time: 4.32 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 2 Compound 2:3-[[trans-4-(Diethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using diethylphosphinic chloride in place of dimethylphosphinicchloride. MS (m/z): 550.3 [M+H]⁻; HPLC retention time: 4.77 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 3 Compound 3:5-(3,3-Dimethyl-but-1-ynyl)-3-{(trans-4-methyl-cyclohexanecarbonyl)-[trans-4-(methyl-phenyl-phosphinoyloxy)-cyclohexyl]-amino}thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using methylphenylphosphinic chloride in place of dimethylphosphinicchloride: MS (m/z): 584.3 [M+H]⁻; HPLC retention time: 4.99 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 4 Compound 4:5-(3,3-Dimethyl-but-1-ynyl)-3-[[(trans-4-(diphenyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using diphenylphosphinic chloride in place of dimethylphosphinicchloride: MS (m/z): 646.3 [M+H]⁻; HPLC retention time: 5.47 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 5 Compound 5:5-(3,3-Dimethyl-but-1-ynyl)-3-[[cis-4-(dimethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using5-(3,3-Dimethyl-but-1-ynyl)-3-[(cis-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid. MS (m/z): 522.2 [M+H]⁻; HPLC retention time: 4.31 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 6 Compound 6:5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-(dimethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-trifluoromethyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-trifluoromethyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid. MS (m/z): 576.2 [M+H]⁻; HPLC retention time: 4.15 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 7 Compound 7:3-[[trans-4-(Dimethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-{4[(thiazole-4-carbonyl)-amino]-phenyl}-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using3-[(trans-4-Hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-{4-[(thiazole-4-carbonyl)-amino}-phenyl]-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid: MS (m/z): 644.2 [M+H]⁻; HPLC retention time: 3.62 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 8 Compound 8:3-[[trans-4-(Dimethyl-phosphinoyloxy)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(4-pyrazolo[1,5-a]pyrimidin-2-yl-phenyl)-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using3-[(trans-4-Hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(4-pyrazolo[1,5-a]pyrimidin-2-yl-phenyl)-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]thiophene-2-carboxylicacid: MS (m/z): 635.3 [M+H]⁻; HPLC retention time: 3.76 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 9 Compound 9:5-(3,3-Dimethyl-but-1-ynyl)-3-[[4-(1-methoxy-1-oxo-1λ⁵-phosphinan-4-ylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

Synthesis of 3-(N-(4-aminocyclohexyl)-(trans-4-ethylcyclohexanecarboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylic acidhydrochloride

To the mixture of methyl5-(3,3-dimethylbut-1-ynyl)-3-(4-methyl-N-(4-oxocyclohexyl)cyclohexanecarboxamido)thiophene-2-carboxylate(440 mg, 1 mmol), ammonium acetate (745 mg, 10 mmol) and sodium sulfate(250 mg) in methanol was added sodium triacetoxyborohydride (424 mg, 2mmol). The reaction mixture was stirred at room temperature for 2 h.Sodium sulfate was removed by filtration and the filtrate was dilutedwith ethyl acetate, washed with saturated sodium bicarbonate and brine.The organic layer was dried over magnesium sulfate, filtered andconcentrated. The residue was purified by HPLC (Gemini column; 5%acetonitrile:water, 5 min; 5-100% acetonitrile:water, 18 min; 100%acetonitrile, 6 min; both solvents containing 0.1% trifluoroaceticacid). The combined fractions were concentrated, basified and extractedwith EtOAc. The organic layer was dried over magnesium sulfate, filteredand concentrated to give title compound (230 mg): MS (m/z): 459.3 [M+H]

Methyl 3-(N-(4-aminocyclohexyl)-(trans-4-methylcyclohexanecarboxamide)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylate (138 mg,0.3 mmol) was dissolved in 4 mL of methanol and 2 mL of THF, and to thesolution was added 2 mL of 1N LiOH. The reaction mixture was stirred atroom temperature for 2 h. Volatiles were removed under vacuum and theresidue was acidified to pH3 using 0.5 N HCl and then partitionedbetween ethyl acetate and water. The organic layer was dried overmagnesium sulfate, filtered and concentrated to give title compound (126mg): MS (m/z): 445.2 [M+H]

Synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-[[4-(1-methoxy-1-oxo-1λ⁵-phosphinan-4-ylamino)-cyclohexyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

3-(N-(4-aminocyclohexyl)-4-methylcyclohexanecarboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylicacid hydrochloride (40 mg, 0.083 mmol) and1-methoxy-1-oxo-1λ⁵-phosphinan-4-one (27 mg, 0.166 mmol) were dissolvedin 2 mL of DMF. Sodium triacetoxyborohydride (35 mg, 0.166 mmol) wasadded. The reaction mixture was allowed to stir at room temperature over18 hours, and then diluted with EtOAc, washed with saturated sodiumbicarbonate and brine. The organic layer was dried over magnesiumsulfate, filtered and concentrated. The residue was purified by HPLC(Gemini column; 5% acetonitrile:water, 5 min; 5-100% acetonitrile:water,18 min; 100% acetonitrile, 6 min; both solvents containing 0.1%trifluoroacetic acid) to give title compound (38.6 mg): MS (m/z): 591.3[M+H]⁻; HPLC retention time: 3.28 min (2-98% acetonitrile:water with0.05% trifluoroacetic acid).

Example 10 Compound 10:5-(3,3-Dimethyl-but-1-ynyl)-3-[[1-(1-methoxy-1-oxo-1λ⁵-phosphinan-4-yl)-piperidin-4-yl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 9,using5-(3,3-dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-piperidin-4-yl-amino]-thiophene-2-carboxylicacid in place of3-[(4-Amino-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid: MS (m/z): 577.1 [M+H]⁻; HPLC retention time: 3.46 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 11 Compound 11:5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

Step 1: Synthesis of Methyl5-(3,3-Dimethyl-but-1-ynyl)-3-(1-methoxy-1-oxo-1λ⁵phosphinan-4-ylamino)-thiophene-2-carboxylate

A solution of methyl3-amino-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylate (474 mg, 2mmol) and 1-Methoxy-1-oxo-1λ⁵ phosphinan-4-one (390 mg, 2.4 mmol) in THF(1 mL) and DMF (1 mL) was treated with dibutyltin dichloride (60 mg, 0.2mmol). After 5 min, phenylsilane (272 μL, 2.2 mmol) was added, and themixture was stirred for one hour at room temperature and thenpartitioned between water and ethyl acetate. The aqueous phase wasextracted with ethyl acetate. The combined organic layers were washedwith 5% LiCl and brine, dried over magnesium sulfate, filtered andconcentrated. The residue was purified by silica gel columnchromatography using EtOAc:Hexanes as eluent to provide 650 mg (85%) oftitle compound: MS (m/z) 384.0 [M+H]⁻; HPLC retention time: 4.463 min(2-98% acetonitrile:water with 0.05% trifluoroacetic acid).

Step 2: Synthesis of methyl5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵-phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylate

Methyl5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵phosphinan-4-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylate(420 mg, 1.1 mmol) was dissolved in 2 mL of THF and cooled to −78° C.with a dry ice-acetone bath. A solution of KHMDS (2.6 mL, 0.5 M intoluene) was added slowly, and the mixture was stirred for 5 min.Trans-4-methylcyclohexanecarbonyl chloride (210 mg, 1.3 mmol) was addeddropwise and stirring was continued at −78° C. for a further 5 min. Themixture was then allowed to warm to room temperature. The reactionmixture was cooled to 0° C. and quenched with saturated aqueous NH₄Cl,and then partitioned between water and ethyl acetate. The aqueous phasewas extracted with ethyl acetate. The combined organic layers werewashed with brine, dried over magnesium sulfate, filtered andconcentrated. The residue was purified by silica gel columnchromatography using 10% MeOH in EtOAc: Hexanes as eluent and thenfurther purified by HPLC (Gemini column; 5% acetonitrile:water, 5 min;5-100% acetonitrile:water, 18 min; 100% acetonitrile, 6 min; bothsolvents containing 0.1% trifluoroacetic acid), resulting in the titlecompound (37 mg) and recovered starting material (210 mg). MS (m/z):508.2 [M+H]⁺; HPLC retention time: 4.949 min (2-98% acetonitrile:waterwith 0.05% trifluoroacetic acid).

Step 3: Synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵-phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵-phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylate(36 mg, 0.07 mmol) was dissolved in 1 mL of methanol and 1 mL of THF,and to the solution was added 0.2 mL of 1N LiOH. The reaction mixturewas stirred at room temperature for 5 hours, and then cooled to 0° C.and neutralized with 0.4 mL of 0.5 N HCl. After removal of solvent, theresidue was purified by HPLC (Gemini column; 5% acetonitrile:water, 5min; 5-100% acetonitrile:water, 18 min; 100% acetonitrile, 6 min; bothsolvents containing 0.1% trifluoroacetic acid), resulting in 20 mg (57%)of the title compound. MS (m/z): 494.2 [M+H]⁻; HPLC retention time:4.144 min (2-98% acetonitrile:water with 0.05% trifluoroacetic acid).

Example 12 Compound 12:5-(3,3-Dimethyl-but-1-ynyl)-3[(1-hydroxy-1-oxo-1λ⁵phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

5-(3,3-Dimethyl-but-1-ynyl)-3-[(1-methoxy-1-oxo-1λ⁵-phosphinan-4-yl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (10 mg, 0.02 mmol) was dissolved in 1 mL of CH₂Cl₂, and to thesolution was added TMSBr (26 μL, 0.2 mmol). The reaction mixture wasstirred at room temperature for 40 minutes. Another portion of TMSBr (26μL, 0.2 mmol) was added and the mixture was stirred at room temperaturefor 5 hours before cooling to 0° C. and quenching through the additionof methanol (0.5 ml). The solvent was removed under vacuum and theresidue was purified by HPLC (Gemini column; 5% acetonitrile:water, 5min; 5-100% acetonitrile:water, 18 min; 100% acetonitrile, 6 min; bothsolvents containing 0.1% trifluoroacetic acid). This resulted in 6 mg(62%) of the title compound. MS (m/z): 480.2 [M+H]⁺; HPLC retentiontime: 3.831 min (2-98% acetonitrile:water with 0.05% trifluoroaceticacid).

Example 13 Compound 13:5-(3,3-dimethylbut-1-ynyl)-3-((1r,4r)-N-(4-ethoxy(methyl)phosphoryl)-4-hydroxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylicacid

5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylicacid (100 mg, 0.23 mmol), ethyl methylphosphinate (49 mg, 0.46 mmol) andtriethylamine (96 μL, 0.69 mmol) were heated with stirring at 100° C.for 2 hours and then cooled to room temperature. The solvent was removedunder vacuum and the residue purified by HPLC (Gemini column; 5%acetonitrile:water, 5 min; 5-100% acetonitrile:water, 18 min; 100%acetonitrile, 6 min; both solvents containing 0.1% trifluoroaceticacid). This resulted in 35 mg of the title compound (as a mixture ofcis- and trans-isomers): MS (m/z): 552.2 [M−H]⁺; HPLC retention time:4.111 min and 4.211 min (2-98% acetonitrile:water with 0.05%trifluoroacetic acid)

Example 14 Compound 14:5-(3,3-dimethylbut-1-ynyl)-3-((1r,4r)-N-(4-(ethoxy(methyl)phosphoryl)-4-hydroxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylicacid

Step 1: Synthesis of methyl5-(3,3-dimethylbut-1-ynyl)-3-(N-(4-(ethoxy(methyl)phosphoryl)-4-hydroxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylate

5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-methyl-cyclohexanecarbonyl)-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylicacid methyl ester (500 mg, 1.1 mmol), ethyl methylphosphinate (238 mg,2.2 mmol) and triethylamine (306 μL, 2.2 mmol) were heated at 100° C.for 2 hours with stirring and then cooled down to room temperature.Further portions of ethyl methylphosphinate (119 mg, 1.1 mmol) andtriethylamine (153 μL, 1.1 mmol) were added and heating at 100° C. wascontinued for 5 hours. The reaction mixture was cooled to roomtemperature and the solvent was removed under vacuum. The residue waspurified by HPLC (Gemini column; 5% acetonitrile:water, 5 min; 5-100%acetonitrile:water, 18 min; 100% acetonitrile, 6 min; both solventscontaining 0.1% trifluoroacetic acid). This resulted in 380 mg of thetitle compound (mixture of cis- and trans-isomers): MS (m/z): 566.2[M+H]⁺; HPLC retention time: 4.860 min and 4.955 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Step 2: Synthesis of methyl5-(3,3-dimethylbut-1-ynyl)-3-(N-(4-(ethoxy(methyl)phosphoryl)-4-methoxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylate

Methyl5-(3,3-dimethylbut-1-ynyl)-3-(N-(4-(ethoxy(methyl)phosphoryl)-4-hydroxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylate(85 mg, 0.15 mmol, mixture of cis- and trans-isomers) and iodomethane(47 μL, 0.75 mmol) were dissolved in 0.5 mL of DMF and cooled to 0° C. Asuspension of NaH (12 mg, 0.30 mmol, 60% dispersion in mineral oil) in0.5 mL of DMF was added slowly and then the reaction mixture was allowedto stir at room temperature for 20 min. The reaction was neutralized at0° C. by the addition of saturated aqueous NH₄Cl and then partitionedbetween water and ethyl acetate. The aqueous phase was extracted withethyl acetate. The combined organic layers were washed with 5% LiCl andbrine, dried over magnesium sulfate, filtered and concentrated. Theresidue was purified by HPLC (Gemini column; 5% acetonitrile:water, 5min; 5-100% acetonitrile:water, 18 min; 100% acetonitrile, 6 min; bothsolvents containing 0.1% trifluoroacetic acid). This resulted in 60 mgof the title compound. MS (m/z): 580.3 [M+H]⁺; HPLC retention time:5.434 min and 5.700 min (2-98% acetonitrile:water with 0.05%trifluoroacetic acid).

Step 3: Synthesis of5-(3,3-dimethylbut-1-ynyl)-3-(N-(4-(ethoxy(methyl)phosphoryl)-4-methoxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylicacid

Methyl5-(3,3-dimethylbut-1-ynyl)-3-(N-(4-(ethoxy(methyl)phosphoryl)-4-methoxycyclohexyl)-(trans-4-methylcyclohexanecarboxamido)thiophene-2-carboxylate(60 mg, 0.1 mmol) was dissolved in 0.5 mL of methanol and 0.5 mL of THF,and to the solution was added 0.3 mL of 1N LiOH. The reaction mixturewas allowed to stir at room temperature for 3 hours, cooled to 0° C. andneutralized with 0.5 N HCl. The solvent was removed under vacuum. Theresidue was purified by HPLC (Gemini column; 5% acetonitrile:water, 5min; 5-100% acetonitrile:water, 18 min; 100% acetonitrile, 6 min; bothsolvents containing 0.1% trifluoroacetic acid). This resulted in 9 mg ofone isomer of the title compound (MS (m/z): 566.3 [M+H]⁺; HPLC retentiontime: 4.544 min (2-98% acetonitrile:water with 0.05% trifluoroaceticacid)) and 9 mg of the other isomer (MS (m/z): 566.3 [M+H]⁺; HPLCretention time: 4.751 min (2-98% acetonitrile:water with 0.05%trifluoroacetic acid)).

Example 15 Compound 15:5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(dimethyl-phosphinoyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

Step 1: Synthesis of3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethylamino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester

The title compound was synthesized in a manner analogous to Example 11step 1, using (tent-Butyl-dimethyl-silanyloxy)-acetaldehyde in place of1-Methoxy-1-oxo-1λ⁵phosphinan-4-one; reaction time was 24 hours.

Step 2: Synthesis of3-[[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester

The title compound was synthesized in a manner analogous to Example 11step 2, using3-[2-(tert-Butyl-dimethyl-silanyloxy)-ethylamino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester in place of methyl5-(3,3-Dimethyl-but-1-ynyl)-3-(1-methoxy-1-oxo-1λ⁵phosphinan-4-ylamino)-thiophene-2-carboxylate:MS (m/z): 520.3 [M+H].

Step 3: Synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-[(2-hydroxy-ethyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

3-[[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester (450 mg, 0.86 mmol) in a 1:1:1 mixture ofTHF:MeOH:0.5N lithium hydroxide (18 mL) was stirred at room temperaturefor 24 hours. The reaction mixture was acidified with 10% HCl_((ac))then partitioned between ethyl acetate and water. The organic phase wasdried over sodium sulfate then concentrated to dryness. The residue waspurified by silica gel chromatography (10-50% ethyl acetate:hexanes),resulting in 273 mg of3-[[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid.

3-[[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid (273 mg) in THF (4 mL) was treated withtriethylaminetrihydrofluoride (0.5 mL). After 1 hour, volatiles wereremoved under vacuum and the residue was purified by silica gelchromatography (0-10% Methanol:DCM) to provide 83 mg of5-(3,3-Dimethyl-but-1-ynyl)-3-[(2-hydroxy-ethyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid.

Step 4: Synthesis of3-[[2-(Dimethyl-phosphinoyloxy)-ethyl]-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 1,using5-(3,3-Dimethyl-but-1-ynyl)-3-[(2-hydroxy-ethyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid: MS (m/z): 468.2 [M+H]⁻; HPLC retention time: 4.144 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 16 Compound 16:3-[[2-(Diethyl-phosphinoyloxy)-ethyl]-(trans-4-methyl-cyclohexariecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid

The title compound was synthesized in a manner analogous to Example 15,using diethylphosphinic chloride in place of dimethylphosphinicchloride: MS (m/z): 496.2 [M+]⁻; HPLC retention time: 2.70 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 17 Compound 17:5-(3,3-Dimethyl-but-1-ynyl)-3-[[trans-4-(dimethyl-phosphinoyloxy)-cyclobutyl]-(cis-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

A mixture of 3-amino-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester, hydrochloride salt (750 mg, 2.74 mmol) and3-benzyloxy-cyclobutanone (2.5 g, 14.3 mmol) in DCM (40 mL) was treatedwith sodium triacetoxyborohydride (3.34 g, 15.8 mmol) portionwise. Themixture was stirred at room temperature for 3 hours. The solution wasdiluted with ethyl acetate, washed sequentially with saturated aqueoussodium bicarbonate, water, and brine. The organic phase was dried overmagnesium sulfate, filtered and concentrated. The residue was purifiedby silica gel chromatography (0-10% ethyl acetate:hexanes). Thisresulted in 1.26 g (quant. yield) of3-(3-benzyloxy-cyclobutylamino)-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester as a mixture of cis- and trans-isomers.

A mixture of3-(3-Benzyloxy-cyclobutylamino)-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester (1.26 g, 3.17 mmol) and N,N-diisopropylethylamine (2.4mL, 13.8 mmol) in DCE (8 mL) was treated withtrans-4-methyl-cyclohexanecarbonyl chloride and heated to 100° C. for 5hours. The reaction mixture was cooled to room temperature, adsorbedonto silica, and purified by silica gel chromatography (0-30% ethylacetate:hexanes). This resulted in 590 mg (36% yield) of cis-cyclobutylisomer(cis-3-[(3-benzyloxy-cyclobutyl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester) and 230 mg (14% yield) of the trans-cyclobutyl isomer(trans-3-[(3-benzyloxy-cyclobutyl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester).

3-[(cis-3-benzyloxy-cyclobutyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-5-(3,3-dimethyl-but-1-ynyl)-thiophene-2-carboxylicacid methyl ester (290 mg, 0.55 mmol) in DCM (2 mL) at 0° C. was treatedwith boron tribromide (0.85 mL, 1.0 M solution in DCM). The mixture wasstirred at 0° C. for 15 minutes and quenched with the addition of silicagel. Volatiles were evaporated under reduced pressure and the reactionmixture was purified by silica gel chromatography (0-100% ethylacetate:hexanes). This resulted in 210 mg (88% yield) of5-(3,3-Dimethyl-but-1-ynyl)-3-[(cis-3-hydroxy-cyclobutyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester.

5-(3,3-Dimethyl-but-1-ynyl)-3-[(cis-3-hydroxy-cyclobutyl)-(trans-4-methyl-cyclohexancarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester (400 mg, 0.93 mmol) in a 3:2:1 mixture ofTHF:MeOH:water (20 mL) was treated with lithium hydroxide (4.5 mL, 1.0 Maqueous solution) and heated to 60° C. for 2 hours. The organicvolatiles were evaporated under reduced pressure and remaining solutionwas acidified with 10% HCl_((aq)). A white precipitate was collected byvacuum filtration, washed with water, and dried to afford 290 mg (74%yield) of5-(3,3-Dimethyl-but-1-ynyl)-3-[(cis-3-hydroxy-cyclobutyl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid as a white powder. MS (m/z): 415.8 [M−H]⁻; HPLC retention time:4.10 min (2-98% acetonitrile:water with 0.05% trifluoroacetic acid).

The title compound was synthesized in a manner analogous to Example 1,using5-(3,3-dimethyl-but-1-ynyl)-3-[(cis-3-hydroxy-cyclobutyl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid in place of5-(3,3-Dimethyl-but-1-ynyl)-3-[(trans-4-hydroxy-cyclohexyl)-(trans-4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid. MS (m/z): 495.1 [M−H]⁻; HPLC retention time: 4.57 min (2-98%acetonitrile:water with 0.05% trifluoroacetic acid).

Example 18 Compound 18:5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(methoxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

PtO₂ (400 mg) was slurried with EtOH (20 mL). To the slurry was added asolution of cyanomethyl-phosphoric acid diethyl ester (4 g, 22.58 mmol)in EtOH (40 mL), followed by 12N HCl (2 mL). The reaction vessel'satmosphere was exchanged with H₂ and the reaction was stirred at roomtemperature until complete, as determined by TLC (eluent: 100% EtOAc).The reaction was then filtered through Supercell NF. The filtrate wasconcentrated to yield the HCl salt of (2-amino-ethyl)-phosphoric aciddiethyl ester as a white solid (4.55, 93%).

The HCl salt of (2-amino-ethyl)-phosphonic acid diethyl ester (4.5 g,20.7 mmol) was dissolved in CH₂Cl₂ (100 mL). To this solution was addedCBzCl (3.2 mL, 22.7 mmol), followed by iPr₂NEt (14.4 mL, 82.8 mmol). Thereaction was stirred at room temperature until complete, as determinedby TLC (eluent: 100% EtOAc). The reaction was quenched by the additionof saturated NaHCO₃. The mixture was then partitioned between CH₂Cl₂ andH₂O. The layers were separated and the organic layer was washed withbrine and then dried over Na₂SO₄. The drying agent was removed by vacuumfiltration and (2-benzyloxycarbonylamino-ethyl)-phosphoric acid diethylester was isolated from the concentrated filtrate by silica gel columnchromatography (EtOAc/hexanes) (5.98 g, 92%).

(2-Benzyloxycarbonylamino-ethyl)-phosphonic acid diethyl ester (5.77 g,18.3 mmol) was dissolved in pyridine (61 mL). To this solution was addedNaI (13.7 g, 92 mmol). The mixture was heated at 115° C. overnight. Thenext day the reaction was determined, by LC/MS, to be largely complete.The reaction was cooled to room temperature and concentrated in vacuo.The residue was dissolved in H₂O and extracted with Et₂O. The water wasthen cooled in an ice bath and the adjusted to pH 2 with 2N HCl. Thewater was then extracted with CH₂Cl₂ and the combined organic layerswere washed with brine and dried over Na₂SO4. The drying agent wasremoved by vacuum filtration and the filtrate was concentrated to yield(2-benzyloxycarbonylamino-ethyl)-phosphonic acid monoethyl ester (4.68g, 89%) as an oil that turned to a waxy solid under high vacuum.

Oxaloyl chloride (2.88 mL, 33 mmol) was dissolved in CH₂Cl₂ (42 mL). DMF(284 □L) was added to this solution in a drop-wise manner. After 15minutes a solution of (2-benzyloxycarbonylamino-ethyl)-phosphoric acidmonoethyl ester (4.75 g, 16.5 mmol) in solution with CH₂Cl₂ (42 mL) wasadded drop-wise over 20 minutes at room temperature. The reaction wasstirred at room temperature for 40 minutes. The reaction was determinedto be complete by the absence of starting material in a ³¹P NMR spectrumof the crude reaction mixture. The reaction was concentrated in vacuo toyield an orange-brown oil that was placed under high vacuum overnight.The oil was then dissolved in THF (80 mL) and cooled to −30° C., asdetermined by an internal thermometer. A 1M solution of MeMgBr in amixture of toluene and THF was then slowly added to the reaction whilekeeping the internal temperature at about −30° C. The mixture wasstirred at −30° C. for 45 minutes and then another 1 equivalent ofMeMgBr was added. After stirring at −30° C. for another 45 minutes thereaction was quenched by the addition of saturated NH₄Cl. The reactionwas warmed to room temperature and then partitioned between EtOAc andH₂O. The layers were separated and the organic layer was washed withbrine and dried over Na₂SO₄.(2-Benzyloxycarbonylamino-ethyl)-methyl-phosphinic acid ethyl ester wasisolated from the concentrated filtrate (1.33 g, 28%) by silica gelcolumn chromatography (EtOAc/hexanes then MeOH/CH₂Cl₂) as a yellowcrystalline solid.

(2-Benzyloxycarbonylamino-ethyl)-methyl-phosphinic acid ethyl ester (1.3g, 4.56 mmol) and 10% Pd/C (485 mg) were combined in a flask and EtOH(46 mL) was added. The reaction flask atmosphere was exchanged for H₂and the reaction was run at room temperature until complete, asdetermined by TLC (eluent: 100% EtOAc). The reaction was then filteredthrough Supercell NF and the filtrate was concentrated to yield(2-amino-ethyl)-methyl-phosphinic acid ethyl ester (615 mg, 90%) as ayellow oil in which white crystals began to form under high vacuum.

5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid methylester (669 mg, 1.92 mmol), Pd(OAc)₂ (64 mg, 0.288 mmol), BINAP (179 mg,0.288 mmol), and Cs₂CO₃ (1.56 g, 4.8 mmol) were combined in a sealedtube. A solution of (2-amino-ethyl)-methyl-phosphinic acid ethyl ester(578 mg, 3.85 mmol) in toluene (14 mL), which had been degassed bybubbling argon through it for 30 minutes, was added. The sealed tube waspurged with argon and then sealed. The tube was placed in a 120° C. oilbath. The reaction was stirred overnight, then cooled to roomtemperature and filtered through filter paper. The residue was stirredvigorously with EtOAc and then the EtOAc was passed through filterpaper. The combined filtrates were concentrated and5-(3,3-Dimethyl-but-1-ynyl)-3-[2-(ethoxy-methyl-phosphinoyl)-ethylamino]-thiophene-2-carboxylicacid methyl ester (466 mg, 66%) was isolated by silica gel columnchromatography (MeOH/EtOAc) as a green crystalline solid.

5-(3,3-Dimethyl-but-1-ynyl)-3-[2-(ethoxy-methyl-phosphinoyl)-ethylamino]-thiophene-2-carboxylicacid methyl ester (233 mg, 0.629 mmol) was dissolved in pyridine (4 mL)and then 4-methyl-cyclohexanecarbonyl chloride (506 mg, 3.15 mmol) wasadded. The reaction was stirred for 7 hours at 100° C., the heat to theoil bath was shut off and the reaction was allowed to stir for a further9 hours. The reaction was diluted with EtOAc and extracted with H₂O. Theorganic phase was then extracted with brine and dried over Na₂SO₄. Thedrying agent was removed by vacuum filtration and the filtrate wasconcentrated.5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(ethoxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester (251 mg, 78%) was isolated from the residue by silicagel column chromatography.

5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(ethoxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester (251 mg, 0.51 mmol) was dissolved in THF (7.3 mL) andMeOH (7.3 mL). A 1N solution of LiOH in H₂O (1.52 mL, 1.52 mmol) wasadded to the reaction drop-wise at room temperature. The reaction wasrun at room temperature until complete, as determined by LC/MS. Thereaction was cooled in an ice bath and then acidified to pH 3 with 1NHCl. The entire mixture was then evaporated in vacuo and stored underhigh vacuum overnight.5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(ethoxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (168 mg, 69%) was isolated from the residue by reverse phase HPLC.MS (m/z): 481.93 [M+H]+; ³¹P NMR (161.9 MHz, CDCl₃): d 56.771, 55.816.

The synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid methylester, required in the above route, is illustrated below:

To a solution of 5-(3,3-Dimethyl-but-1-ynyl)-thiophene-2-carboxylic acid(6.2 g, 30 mmol; see patent application U.S. Pat. No. 5,861,421) in THF(100 mL) was added a solution of nBuLi (2.0 M in pentane, 33 mL, 66mmol) via an addition funnel at −78° C. After addition, the reaction wasstirred at −78° C. for 1 h. A solution of I₂ (7.7 g, 30 mmol) in THF(100 mL) was added slowly (ca. 15 min) to the flask. After a further 10mins, the reaction was quenched with 1 N HCl (50 mL) and warmed to roomtemperature. The volatiles were removed in vacuo and the residue wasdissolved in ether (500 mL). The organic solution was washed with 1 MNa₂S₂O₃ (100 mL×2), brine (100 mL) and dried over Na₂SO₄. Afterconcentrated in vacuo, the residue was purified by silica gelchromatography (EtOAc/hexanes) to give5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (5.9 g,65%) as a white solid.

To a solution of5-(3,3-Dimethyl-but-1-ynyl)-3-iodo-thiophene-2-carboxylic acid (1.0 g,3.0 mmol) and DMF (20 μL) in dry dichloromethane (10 mL) was addedoxalyl chloride (508 μL, 6.0 mmol) at room temperature. After stirringat room temperature for 90 min, the reaction was concentrated in vacuoto remove volatiles. The residue was dissolved in pyridine (5 mL) andmethanol (5 mL) and stirred for 2 h. The volatiles were removed in vacuoand the residue was participated between ether (150 mL) and saturatedNH₄Cl solution (50 mL). The organic layer was washed with saturatedNH₄Cl solution (50 mL) and dried over Na₂SO₄. After concentration invacuo, the residue was purified by silica gel chromatography(EtOAc/hexanes) to give the desired product (835 mg, 80%).

Example 19 Compound 19:5-(3,3-dimethyl-but-1-ynyl)-3-[[2-(hydroxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(ethoxy-methyl-phosohinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (108 mg, 0.225 mmol) was dissolved in acetonitrile (3 mL). Thissolution was cooled in an ice bath and TMSI (160 μL, 1.12 mmol) wasadded dropwise. The reaction turned yellow and some precipitate wasformed. After approximately 1 minute 2,6-lutidine (156 μL, 1.35 mmol)was added, resulting in a clear solution. The ice bath was removed andthe reaction was stirred at room temperature for 2 hours. AdditionalTMSI (50 μL) and 2,6-lutidine (50 μL) were added. After 45 minutes thereaction was determined by LC/MS to be complete. The reaction was cooledin an ice bath and additional 2,6-lutidine (100 μL) was added, followedby MeOH (2 mL). The reaction was warmed to room temperature andconcentrated under vacuum.5-(3,3-Dimethyl-but-1-ynyl)-3-[[2-(hydroxy-methyl-phosphinoyl)-ethyl]-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (37 mg, 36%) was isolated from the residue by reverse phase HPLC.MS (m/z): 453.93 [M+H]+; ³¹P NMR (161.9 MHz, CD₃OD): δ 49.909.

Example 24 Compound 24:3-(1,4-N-(4-(2-(diethoxyphosphoryl)ethylamino)cyclohexyl)-4-methylcyclohexanecarboxamido)-5-(3,3-dimethylbut-1-ynyl)thiophene-2-carboxylicacid

Compound 24 was prepared by reductive amination using the same method asdescribed for Example 9, Step 1.

-   MS=609.24-   Retention time: 2.18 min-   LC: Thermo Finnigan PDA Detector-   MS: Thermo Scientific LCQ Fleet-   Column: Phenomenex Gemini-nx 3u C18 110A 30×3 mm-   Solvents: Acetonitrile with 0.1% trifluoroacetic acid, Water with    0.1% trifluoroacetic acid-   Gradient: 0 min-3.1 min 2%-100% ACN, 3.1 min-3.75 min 100% ACN

Synthesis of (1S,6S)-4,6-dimethyl-cyclohex-3-enecarboxylic acid

4S-benzyl-3-(4,6S-dimethyl-cyclohex-3-ene-1S-carbonyl)-oxazolidin-2-one,prepared in a method similar to that described in J. Am. Chem. Soc.110(4), 1988, 1238-1256, was dissolved in THF (1000 mL) and H₂O (350mL). The solution was cooled in an ice bath and 30% H₂O₂ (36 mL, 354mmol) was slowly added followed by LiOH*H₂O (9.90 g, 263 mmol) in oneportion. The reaction was allowed to slowly warm to rt and was stirredfor 16 h. The reaction was then cooled in an ice bath. Na₂SO₃ (60 g, 472mmol) was dissolved H₂O (400 mL) and very slowly added to the cooledreaction mixture. The solution was stirred for 1 h, and the layers wereseparated. The organic layer was concentrated under reduced pressure.The aqueous layer was added back to the organic concentrate and waswashed with CH₂Cl₂ (2×500 mL). The pH of the aqueous layer was adjustedto 2 through a slow addition of conc. HCl. The aqueous layer wasextracted with EtOAc (4×300 mL) and dried over Na₂SO₄. The ethyl acetateextracts were combined and concentrated under reduced pressureco-evaporating with hexanes to afford(1S,65)-4,6-dimethyl-cyclohex-3-enecarboxylic acid (14.14 g, 78%) as awhite solid.

Synthesis of (1R,6R)-4,6-dimethyl-cyclohex-3-enecarboxylic acid

4,6R-Dimethyl-cyclohex-3-ene-1R-carboxylic acid was prepared in asimilar manner to 4,6S-dimethyl-cyclohex-3-ene-1S-carboxylic acid, usingthe chiral auxiliary 4R-benzyl-oxazolidin-2-one.

Synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-[(1S,6S)-(4,6-dimethyl-cyclohex-3-enecarbonyl)-(4-hydroxy-cyclohexyl)-amino]-thiophene-2-carboxylicacid

(1S,6S)-4,6-dimethyl-cyclohex-3-ene-carboxylic acid (3.04 g, 19.7 mmol)was dissolved in CH₂Cl₂ (30 mL) and DMF (20 μL) was added. The solutionwas cooled to 0° C. and (COCl)₂ (3.7 mL, 39 mmol) was added slowly. Thereaction was stirred in an ice bath for 2 hours and then concentrated.The residue was taken up in hexanes and concentrated; this hexanescoevaporation was repeated once more. To the residue was added[5-(3,3-Dimethyl-but-1-ynyl)-thiophen-3-yl]-(1,4-dioxa-spiro[4.5]dec-8-yl)-amine(4.16 g, 13 mmol), diisopropylethylamine (4.5 mL, 26 mmol), and1,2-dichloroethane (40 mL) at 0° C. The solution was warmed to roomtemperature and stirred overnight. The reaction was diluted with CH₂Cl₂,twice washed with saturated NH₄Cl_((aq)), dried over MgSO₄, filtered,concentrated, and purified by silica gel column chromatography, elutingwith a mixture of 0-75% EtOAc/hexanes, to give(1S,6S)-4,6-dimethyl-cyclohex-3-ene-carboxylic acid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(1,4-dioxa-spiro[4.5]dec-8-yl)-amide(5.6 g, 12 mmol) as a single isomer.

(1S,6S)-4,6-Dimethyl-cyclohex-3-ene-carboxylic acid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(1,4-dioxa-spiro[4.5]dec-8-yl)-amide(5.6 g, 12 mmol) was dissolved in THF (70 mL) and treated with 4M HCl(35 mL). The reaction mixture was heated to 45° C. and stirred for 2.5h. THF was removed in vacuo, and the aqueous layer was thrice extractedinto ethyl acetate. The combined organic layers were washed withsaturated NaHCO_(3(aq)), water, and brine, dried over MgSO₄, filtered,and concentrated to give (1S,6S)-4,6-dimethyl-cyclohex-3-ene-carboxylicacid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-oxo-cyclohexyl)-amide(5.05 g, 12 mmol).

(1S,6S)-4,6-Dimethyl-cyclohex-3-ene-carboxylic acid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(4-oxo-cyclohexyl)-amide(2.0 g, 4.9 mmol) in MeOH (100 mL) was treated with sodium borohydride(230 mg, 6.0 mmol) at 0° C. After stirring for 30 min, 4M HCl (6 mL) wasadded and the reaction mixture was twice extracted with ethyl acetate.The combined organic layers washed with saturated NaHCO_(3(aq)), brine,dried over MgSO₄, filtered, and concentrated. Silica gel chromatography(20-60% ethyl acetate/hexanes) gave the desired(1S,6S)-4,6-dimethyl-cyclohex-3-ene-carboxylic acid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-hydroxy-cyclohexyl)-amide(1.74 g, 4.2 mmol).

(1S,6S)-4,6-Dimethyl-cyclohex-3-ene-carboxylic acid[5-(3,3-dimethyl-but-1-ynyl)-thiophen-3-yl]-(trans-4-hydroxy-cyclohexyl)-amide(1.74 g, 4.2 mmol) in THF (50 mL) was cooled to −78° C. and treated withlithium diisopropylamine (8.4 mL, 2.0M in heptane/THF/PhEt, 16.8 mmol)and allowed to warm to 0° C. over the course of 2 hours. CO₂ wasvigorously bubbled through the reaction solution for 10 minutes. Thereaction was then quenched with the addition of iPrOH, diluted withethyl acetate, washed with saturated NH₄Cl_((aq)), dried over MgSO₄,filtered, and concentrated. Silica gel chromatography (0-100% ethylacetate/dichloromethane) afforded 530 mg (1.2 mmol) of the titlecompound: MS (m/z): 458.1 [M+H]+; HPLC retention time 4.35 min (2-98%acetonitrile: water with 0.05% trifluoroacetic acid).

Example 25 Compound 25:5-(3,3-Dimethylbut-1-ynyl)-3-((1S,6S)-N-((1r,4S)-4-(dimethylphosphoryloxy)cyclohexyl)-4,6-dimethylcyclohex-3-enecarboxamido)thiophene-2-carboxylicacid

Compound 25 may be prepared in the same manner as Compound 1 by startingwith5-(3,3-dimethyl-but-1-ynyl)-3-[(1S,6S)-(4,6-dimethyl-cyclohex-3-enecarbonyl)-(4-hydroxy-cyclohexyl)-amino]-thiophene-2-carboxylicacid.

Synthesis of5-(3,3-Dimethyl-but-1-ynyl)-3-[(1R,6R)-(4,6-dimethyl-cyclohex-3-enecarbonyl)-(4-hydroxy-cyclohexyl)-amino]-thiophene-2-carboxylicacid

By substituting (1R,6R)-4,6-dimethyl-cyclohex-3-enecarboxylic acid for(1S,6S)-4,6-dimethyl-cyclohex-3-enecarboxylic acid in Scheme 1, theabove titled compound may be prepared.

Example 26 Compound 26:5(3,3-Dimethylbut-1-ynyl)-3-((1R,6)-N-((1r,4S)-4-(dimethylphoschoryloxy)cyclohexyl)-4,6-dimethylcyclohex-3-enecarboxamido)thiophene-2-carboxylicacid

Compound 26 may be prepared in the same manner as Compound 25 startingwith5-(3,3-dimethyl-but-1-ynyl)-3-[(1R,6R)-(4,6-dimethyl-cyclohex-3-enecarbonyl)-(4-hydroxy-cyclohexyl)-amino]-thiophene-2-carboxylicacid rather than5-(3,3-dimethyl-but-1-ynyl)-3-[(1S,6S)-(4,6-dimethyl-cyclohex-3-enecarbonyl)-(4-hydroxy-cyclohexyl)-amino]-thiophene-2-carboxylicacid.

Synthesis of (1S)-4-methyl-cyclohex-3-enecarboxylic acid

Acrylic acid 4,4-dimethyl-2-oxo-tetrahydro-furan-3-yl ester (R) (2.92 g,15.9 mmol) in dichloromethane (20 mL) and hexanes (3 mL) was cooled to−10° C. and treated with titanium tetrachloride (2.4 mL, 2.4 M indichloromethane, 2.4 mmol). The red solution was stirred for 15 min andtreated with isoprene (2.4 mL, 23.8 mmol) dropwise over 5 min. Afterstirring for 1.5 h, an additional portion of isoprene (2.4 mL, 23.8mmol) was added and the reaction mixture was stirred at −10 to 0° C. for2.5 h. After cooling to −10° C., the reaction mixture was quenched withammonium chloride (sat. aq.). Water and ethyl acetate:hexanes (1:1) wereadded. The organic layer was separated and the aqueous layer wasextracted again with ethyl acetate:hexanes (1:1). The combined organiclayers were dried over sodium sulfate, filtered and concentrated. Theresidue was purified by flash chromatography (10-40% EtOAc:Hex, 80 gcolumn) to afford 3.35 g (84% yield) of4-methyl-cyclohex-3-(S)-enecarboxylic acid4,4-dimethyl-2-oxo-tetrahydro-furan-3-yl ester as a clear oil.

4-Methyl-cyclohex-3-(S)-enecarboxylic acid4,4-dimethyl-2-oxo-tetrahydro-furan-3-yl ester (3.34 g, 13.2 mmol) inTHF (25 mL), water (2.5 mL) and methanol (2.5 mL) was treated withlithium hydroxide monohydrate (2.8 g, 66.2 mmol) and warmed to 50° C.with stirring. After 1 h, the reaction mixture treated with 1M HCl(about 25 mL). The mixture was extracted with hexanes:ethyl acetate (200mL: 15 mL), dried over sodium sulfate, filtered and concentrated to 2.4g of a white semi-solid. The residue was redissolved inhexanes:dichloromethane (100 mL, 95:5), washed with water, dried oversodium sulfate, filtered and concentrated to 1.68 g (91% yield) of(1S)-4-methyl-cyclohex-3-enecarboxylic acid as a white powder.

Synthesis of methyl5-(3,3-dimethylbut-1-ynyl)-3-((S)—N-((1r,4S)-4-hydroxycyclohexyl)-4-methylcyclohex-3-enecarboxamido)thiophene-2-carboxylate

(1S)-4-Methyl-cyclohex-3-ene-1-carboxylic acid (250 mg, 1.78 mmol),azeotropically dried by evaporation from toluene was dissolved indichloromethane (4 mL) and treated with dimethylformamide (1 drop). Thereaction mixture was cooled to 0° C. and treated dropwise with oxalylchloride (0.42 mL, 4.5 mmol). The reaction mixture was allowed to warmto ambient temperature while stirring for 4 h. The solution wasconcentrated, treated with hexanes and concentrated again to afford(1S)-4-methyl-cyclohex-3-ene-1-carboxylic acid chloride as a lightyellow oil which was used immediately in the next step.

(1S)-4-Methyl-cyclohex-3-ene-1-carboxylic acid chloride (1.8 mmol),dimethyl-but-1-ynyl)-3-(1,4-dioxa-spiro[4.5]dec-8-ylamino)-thiophene-2-carboxylicacid methyl ester (336 mg, 0.89 mmol) and DMAP (217 mg, 1.8 mmol) weredissolved in dichloroethane (2.2 mL), sealed with a cap and heated to80° C. After 2 h, the temperature was increased to 90° C., and thesolution was stirred 16 h. The reaction mixture was further heated to100° C., stirred 24 h and partitioned between water and ethylacetate:hexanes (1:1). The layers were separated and the aqueous layerwas extracted again with ethyl acetate:hexanes (1:1). The combinedorganic layers were dried over sodium sulfate, filtered andconcentrated. Flash chromatography (5% EtOAc:hexanes 5 min then 5-40%EtOAc:hexanes, 20 min, 24 g column) afforded 250 mg (56% yield of thedesired5-(3,3-dimethyl-but-1-ynyl)-3-[(1,4-dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohex-3-enecarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester as a white foam.

5-(3,3-Dimethyl-but-1-ynyl)-3-[(1,4-dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohex-3-enecarbonyl)-amino]-thiophene-2-carboxylicacid methyl ester (240 mg, 0.48 mmol) was dissolved in THF (3.2 mL) andtreated with 4M HCl (1.6 mL, 0.8 mmol). The reaction mixture was heatedto 45° C. and stirred at ambient temperature. After 2 h, methanol wasadded (15-20 drops) and the solution was stirred for 3 h. An additionalportion of 4M HCl (1.6 mL, 0.8 mmol) and methanol (1 mL) were added andthe solution was stirred 16 h at ambient temperature followed by 40° C.for 4 h. The solution was partitioned between water and ethyl acetate.The organic layer was washed with sodium bicarbonate (sat. aq.) andbrine, dried over sodium sulfate and concentrated to 233 mg(quantitative yield) of the desired5-(3,3-dmethyl-but-1-ynyl)-3-[(4-methyl-cyclohex-3-enecarbonyl)-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylicacid methyl ester as a white foam.

5-(3,3-Dimethyl-but-1-ynyl)-3-[(4-methyl-cyclohex-3-enecarbonyl)-(4-oxo-cyclohexyl)-amino]-thiophene-2-carboxylicacid methyl ester (233 mg, 0.51 mmol) in THF (3 mL) and water (0.3 mL)was treated with sodium borohydride (19 mg, 0.48 mmol). After stirringfor 20 min, water was added and the reaction mixture was extract twicewith ethyl acetate. The combined organic layers were dried over sodiumsulfate, filtered and concentrated. Flash chromatography (10%EtOAc:hexanes, 4 min, 10-70% EtOAc:hexanes, 12 min, 12 g column)afforded 130 mg (59% yield) of the desired methyl5-(3,3-dimethylbut-1-ynyl)-3-((S)—N-((1r,4S)-4-hydroxycyclohexyl)-4-methylcyclohex-3-enecarboxamido)thiophene-2-carboxylate.

Example 27 Compound 27: 5-(3,3-dimethylbut-1-ynyl)-3-((S)-N-((1r,4S)-4-(dimethylphosphoryloxy)cyclohexyl)-4-methylcyclohex-3-enecarboxamido)thiophene-2-carboxylicacid

Compound 27 may be prepared by hydrolysis of methyl5-(3,3-dimethylbut-1-ynyl)-3-((S)—N-((1r,4S)-4-hydroxycyclohexyl)-4-methylcyclohex-3-enecarboxamido)thiophene-2-carboxylateto the corresponding acid and conversion of that acid to Compound 27 inthe same manner as for Compound 1.

ADDITIONAL PROPHETIC EXAMPLES

The following additional examples, including enantiomers thereof, may bemade using analogous methods:

wherein R³ is selected from a group in the table below.

Similarly, analogous methods may be used to synthesize compounds havingthe following substitution pattern:

where R³ is selected from

-   —CH₂—P(O)R^(I)R^(II),-   —C(CH₃)₂—P(O)R^(I)R^(II),-   —CH(C₃₋₆ cycloalkyl)-P(O)R^(I)R^(II),-   —CH₂—CH₂—P(O)R^(I)R^(II),-   —C(CH₃)₂—CH₂—P(O)R^(I)R^(II),-   —CH(C₃₋₆cycloalkyl)-CH₂—P(O)R^(I)R^(II), and-   —CH(CH₂R⁺)—CH₂—P(O)R^(I)R^(II), including both the (+) and (−)    configurations about the chiral atom;-   where each R^(I) is independently a variety of substituents, as    herein described, including but not limited to C₁₋₁₂ alkyl, C₁₋₁₂    alkoxy, or —OH;-   where each R^(II) is independently a variety of substituents, as    herein described, including but not limited to C₁₋₁₂ alkyl, C₆₋₁₄    aryl, such as phenyl, or C₈₋₁₆ arylalkyl, such as benzyl; and-   where each R is independently a variety of substituents, as herein    described, including but not limited to C₁₋₁₂ alkyl, or C₁₋₁₂    alkoxy.    General Schemes (for P in R¹)    As in the above general illustrations, the phosphorus-containing    moiety may be introduced into R¹ with the rest of the molecule    already assembled. Alternatively, the bond between the thiophene and    R¹ may conveniently be formed by a variety of aryl-aryl or    aryl-acetylene coupling methods such as those pioneered by Suzuki or    Sonagashira (see Smith, M. B. and March, J. Advanced Organic    Chemistry, 6^(th) edition, Wiley-Interscience, pp 899 and 903    respectively, herein incorporated by reference with regard to such    synthetic teaching), using a 5-halothiophene as illustrated in    Scheme B.

Experimentals

Example 20 Compound 20:3-[(1,4-Dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(ethoxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid

4-(Bromomethyl)benzeneboronic acid pinacol ester (609 mg, 2.05 mmol, 1eq.) was weighed into a round-bottomed flask and purged with N₂.1,4-Dioxane (6 mL) and diethoxymethylphosphine (0.96 mL, 6.15 mmol, 3eq.) were then added, and the solution was stirred at 100° C. for 1hour. The reaction was then concentrated to an oil from which theproduct(methyl-[4-(4,4,5,5-tetramethyl-[1,3,3]dioxaborolan-2-yl)-benzyl]-phosphinicacid ethyl ester) was isolated by column chromatography (SiO₂, 10% MeOHin dichloromethane) as a clear oil (657 mg, 2.028 mmol, 98%). This clearoil was then dissolved in dimethoxyethane (7 mL) and the resultingsolution was added to a 20 mL microwave vial containing intermediate A(3-[(1,4-Dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-iodo-thiophene-2-carboxylicacid methyl ester; see WO 2008/58393 A1; 925 mg, 1.69 mmol, 1 eq.),Pd(PPh₃)₄ (98 mg, 0.0845 mmol, 0.05 eq.), and potassium carbonate (467mg, 3.38 mmol, 2 eq.), and 3.5 mL water. The vial was sealed and heatedby microwave to 120° C. for 10 minutes. The reaction was partitionedbetween ethyl acetate and water. The aqueous layer was washed twice with50 mL ethyl acetate, and the ethyl acetate layers were combined, washedwith brine, dried over magnesium sulfate, filtered, and concentrated toan oil from which intermediate B(3-[(1,4-Dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(ethoxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid methyl ester) was isolated by column chromatography (SiO₂, 9%methanol in ethyl acetate) as a tan solid (1.01 g, 1.65 mmole, 98%).

This material (55 mg, 0.089 mmol, 1 eq.) was dissolved intetrahydrofuran (2 mL) at room temperature. A solution of lithiumhydroxide monohydrate (8 mg, 0.2 mmole, 2.2 eq.) in 1 mL water and 5drops of methanol was then added, and the solution was stirred at roomtemperature for 4 hours. The reaction was neutralized with 0.11 mL 2NHCl and concentrated. The crude material was redissolved in methanol andpurified by reverse phase preparative HPLC to afford3-[(1,4-Dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(ethoxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid (10 mg, 19%). MS (m/z) 604.3 [M+H]⁺HPLC retention time: 3.575 min(5-95% acetonitrile with 0.05% TFA: water with 0.05% TFA).

Example 21 Compound 21:5-[4-(Ethoxy-methyl-phosphinoylmethyl)-phenyl]-3-[(4-hydroxy-cyclohexyl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

3-[(1,4-Dioxa-spiro[4.5]dec-8-yl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(ethoxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid methyl ester (204 mg, 0.331 mmol, 1 eq.) was weighed into a smallflask and dissolved in 3 mL THF. Next, 2 mL of 3.6 N HCl was added andthe reaction was stirred at 40° C. for 1.5 hours. The reaction was thencooled to room temperature and diluted with 40 mL water. The solutionwas then extracted twice with ethyl acetate. The combined ethyl acetatelayers were then washed twice with saturated NaHCO₃, twice with water,and once with brine, dried over sodium sulfate, filtered andconcentrated to afford the crude ketone (176 mg). This material (176 mg,˜0.307 mmol) was dissolved in 5 mL anhydrous THF and cooled to −25° C.using an acetonitrile/dry ice bath. Sodium borohydride (9 mg, 0.230mmol, 0.75 eq.) was added as a solid in two portions. The reaction wasstirred 1.5 hours at −25° C., then was quenched with 3 mL 2 N HCl andwarmed to room temperature. The solution was then extracted twice withethyl acetate. The combined ethyl acetate layers were washed once withbrine, and then dried over sodium sulfate, filtered and concentrated.This crude product (173 mg) was dissolved in 2 mL THF. Stirring wasbegun and 0.5 mL methanol and a solution of lithium hydroxidemonohydrate (25 mg, 0.6 mmol, 2 eq. in 0.5 mL water) were added. Thereaction was stirred at 40° C. for 45 minutes whereupon the reaction wasstopped by the addition of 0.35 mL 2 N HCl. The reaction wasconcentrated, then redissolved in methanol and purified by reverse phasepreparative HPLC to afford5-[4-(Ethoxy-methyl-phosphinoylmethyl)-phenyl]-3-[(4-hydroxy-cyclohexyl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid (11 mg, 6%). MS (m/z) 562.1 [M+H]⁺ HPLC retention time: 3.152 min(5-95% acetonitrile with 0.05% TFA: water with 0.05% TFA).

Example 22 Compound 22:3-[(4-Hydroxy-cyclohexyl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(hydroxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid

Following the reductive step in Example 21, the crude product (50 mg)was dissolved in 1 mL THF and 0.5 mL methanol. 10 N NaOH (0.26 mL, ˜30eq) was added, and the solution was stirred overnight at roomtemperature. The reaction was then neutralized with 1.3 mL 2 N HCl,concentrated, and purified by reverse phase preparative HPLC to afford(3-[(4-Hydroxy-cyclohexyl)-(4-methyl-cyclohexanecarbonyl)-amino]-5-[4-(hydroxy-methyl-phosphinoylmethyl)-phenyl]-thiophene-2-carboxylicacid (20 mg, 11%). MS (m/z) 534.2 [M+H]⁺ HPLC retention time: 2.725 min(5-95% acetonitrile with 0.05% TFA: water with 0.05% TFA).

Example 23 Compound 23:5-[3,3-Dimethyl-5-(tetrahydro-pyran-4-yloxy)-pent-1-ynyl]-3-[(4-(pyridin-2-yloxy)-cyclohexyl)-(4-methyl-cyclohexanecarbonyl)-amino]-thiophene-2-carboxylicacid

A solution of 3,3-dimethyl-pent-4-yn-1-ol (600 mg, 5.36 mmol),3-[[4-(tert-Butyl-dimethyl-silanyloxy)-cyclohexyl]-(4-methyl-cyclohexanecarbonyl)-amino]-5-iodo-thiophene-2-carboxylicacid methyl ester (2767 mg, 4.46 mmol) and triethylamine (10.8 mL, 77.48mmol) in DMF (25 mL) was degassed for 10 minutes with nitrogen.PdCl₂(PPh₃)₂ (314 mg, 0.45 mmol) and CuI (170 mg, 0.90 mmol) was addedand the resulting mixture was degassed with nitrogen for an additional 5minutes after which the reaction was placed in a 80° C. oil bath andheated for 3.5 h. The reaction was cooled and partitioned between ethylacetate and sat. NaHCO₃. The organic layer was separated, washed with 5%LiCl, brine, dried over Na₂SO₄ and concentrated to give a dark brownfoam. Purification by flash column chromatography on silica gel using30% ethyl acetate in hexanes then 5% methanol in ethyl acetate provided3-[[4-(tert-butyl-dimethyl-silanyloxy)-cyclohexyl]-(4-methyl-cyclohexanecarbonyl)-amino]-5-(5-hydroxy-3,3-dimethyl-pent-1-ynyl)-thiophene-2-carboxylicacid methyl ester (2.43 g, 90%) as a pale yellow foam.

After hydrolysis of the3-[[4-(tert-butyl-dimethyl-silanyloxy)-cyclohexyl]-(4-methyl-cyclohexanecarbonyl)-amino]-5-(5-hydroxy-3,3-dimethyl-pent-1-ynyl)-thiophene-2-carboxylicacid methyl ester to the corresponding acid, Compound 23 was obtained byphosphorylated in the same manner as in Example 1.

MS=552; Hplc retention time is 3.319.

PROPHETIC EXAMPLES

The following additional examples, including enantiomers thereof, may bemade using analogous methods:

BIOLOGICAL EXAMPLES

Antiviral Activity

Another aspect of the invention relates to methods of inhibiting viralinfections, comprising the step of treating a sample or subjectsuspected of needing such inhibition with a composition of theinvention.

Within the context of the invention samples suspected of containing avirus include natural or man-made materials such as living organisms;tissue or cell cultures; biological samples such as biological materialsamples (blood, serum, urine, cerebrospinal fluid, tears, sputum,saliva, tissue samples, and the like); laboratory samples; food, water,or air samples; bioproduct samples such as extracts of cells,particularly recombinant cells synthesizing a desired glycoprotein; andthe like. Typically the sample will be suspected of containing anorganism which induces a viral infection, frequently a pathogenicorganism such as a tumor virus. Samples can be contained in any mediumincluding water and organic solvent\water mixtures. Samples includeliving organisms such as humans, and man made materials such as cellcultures.

If desired, the anti-virus activity of a compound of the invention afterapplication of the composition can be observed by any method includingdirect and indirect methods of detecting such activity. Quantitative,qualitative, and semiquantitative methods of determining such activityare all contemplated. Typically one of the screening methods describedabove are applied, however, any other method such as observation of thephysiological properties of a living organism are also applicable.

The antiviral activity of a compound of the invention can be measuredusing standard screening protocols that are known. For example, theantiviral activity of a compound can be measured using the followinggeneral protocols.

Cell-based Flavivirus Immunodetection assay

BHK21 or A549 cells are trypsinized, counted and diluted to 2×10⁵cells/mL in Hams F-12 media (A549 cells) or RPMI-1640 media (BHK21cells) supplemented with 2% fetal bovine serum (FBS) and 1%penicillin/streptomycin. 2×10⁴ cells are dispensed in a clear 96-welltissue culture plates per well and placed at 37° C., 5% CO₂ overnight.On the next day, the cells are infected with viruses at multiplicity ofinfection (MOI) of 0.3 in the presence of varied concentrations of testcompounds for 1 hour at 37° C. and 5% CO₂ for another 48 hours. Thecells are washed once with PBS and fixed with cold methanol for 10 min.After washing twice with PBS, the fixed cells are blocked with PBScontaining 1% FBS and 0.05% Tween-20 for 1 hour at room temperature. Theprimary antibody solution (4G2) is then added at a concentration of 1:20to 1:100 in PBS containing 1% FBS and 0.05% Tween-20 for 3 hours. Thecells are then washed three times with PBS followed by one hourincubation with horseradish peroxidase(HRP)-conjugated anti-mouse IgG(Sigma, 1:2000 dilution). After washing three times with PBS, 50microliters of 3,3′,5,5′-tetramethylbenzidine (TMB) substrate solution(Sigma) is added to each well for two minutes. The reaction is stoppedby addition of 0.5 M sulfuric acid. The plates are read at 450 nmabosorbance for viral load quantification. After measurement, the cellsare washed three times with PBS followed by incubation with propidiumiodide for 5 min. The plate is read in a Tecan Safire™ reader(excitation 537 nm, emission 617 nm) for cell number quantification.Dose response curves are plotted from the mean absorbance versus the logof the concentration of test compounds. The EC₅₀ is calculated bynon-linear regression analysis. A positive control such asN-nonyl-deoxynojirimycin may be used.

Cell-Based Flavivirus Cytopathic Effect Assay

For testing against West Nile virus or Japanese encephalitis virus,BHK21 cells are trypsinized and diluted to a concentration of 4×10⁵cells/mL in RPMI-1640 media supplemented with 2% FBS and 1%penicillin/streptomycin. For testing against dengue virus, Huh7 cellsare trypsinized and diluted to a concentration of 4×10⁵ cells/mL in DMEMmedia supplemented with 5% FBS and 1% penicillin/streptomycin. A 50microliter of cell suspension (2×10⁴ cells) is dispensed per well in a96-well optical bottom PIT polymer-based plates (Nunc). Cells are grownovernight in culture medium at 37° C., 5% CO₂, and then infected withWest Nile virus (e.g. B956 strain) or Japanese encephalitis virus (e.g.Nakayama strain) at MOI=0.3, or with dengue virus (e.g. DEN-2 NGCstrain) at MOI=1, in the presence of different concentrations of testcompounds. The plates containing the virus and the compounds are furtherincubated at 37° C., 5% CO₂ for 72 hours. At the end of incubation, 100microliters of CellTiter-Glo™ reagent is added into each well. Contentsare mixed for 2 minutes on an orbital shaker to induce cell lysis. Theplates are incubated at room temperature for 10 minutes to stabilizeluminescent signal. Lumnescence reading is recorded using a platereader. A positive control such as N-nonyl-deoxynojirimycin may be used.

Antiviral Activity in a Mouse Model of Dengue Infection.

Compounds are tested in vivo in a mouse model of dengue virus infection(Schul at al. J. Infectious Dis. 2007; 195:665-74). Six to ten week oldAG129 mice (B&K Universal Ltd, HII, UK) are housed in individuallyventilated cages. Mice are injected intraperitoneally with 0.4 mL TSV01dengue virus 2 suspension. Blood samples are taken by retro orbitalpuncture under isoflurane anaesthesia. Blood samples are collected intubes containing sodium citrate to a final concentration of 0.4%, andimmediately centrifuged for 3 minutes at 6000 g to obtain plasma. Plasma(20 microliters) is diluted in 780 microliters RPMI-1640 medium and snapfrozen in liquid nitrogen for plaque assay analysis. The remainingplasma is reserved for cytokine and NS1 protein level determination.Mice develop dengue viremia rising over several days, peaking on day 3post-infection.

For testing of antiviral activity, a compound of the invention isdissolved in vehicle fluid, e.g. 10% ethanol, 30% PEG 300 and 60% D5W(5% dextrose in water; or 6N HCl (1.5 eq):1N NaOH (pH adjusted to 3.5):100 mM citrate buffer pH 3.5 (0.9% v/v:2.5% v/v: 96.6% v/v). Thirty six6-10 week old AG129 mice are divided into six groups of six mice each.All mice are infected with dengue virus as described above (day 0).Group 1 is dosed by oral gavage of 200 mL/mouse with 0.2 mg/kg of acompound of the invention twice a day (once early in the morning andonce late in the afternoon) for three consecutive days starting on day 0(first dose just before dengue infection). Groups 2, 3 and 4 are dosedthe same way with 1 mg/kg, 5 mg/kg and 25 mg/kg of the compound,respectively. A positive control may be used, such as(2R,3R,4R,5R)-2-(2-amino-6-hydroxy-purin-9-yl)-5-hydroxymethyl-3-methyl-tetrahydro-furan-3,4-diol,dosed by oral gavage of 200 microliters/mouse the same way as theprevious groups. A further group is treated with only vehicle fluid.

On day 3 post-infection approximately 100 microliter blood samples(anti-coagulated with sodium citrate) are taken from the mice byretro-orbital puncture under isoflurane anaesthesia. Plasma is obtainedfrom each blood sample by centrifugation and snap frozen in liquidnitrogen for plague assay analysis. The collected plasma samples areanalyzed by plague assay as described in Schul at al. Cytokines are alsoanalysed as described by Schul. NS1 protein levels are analysed using aPlatelia™ kit (BioRad Laboratories). An anti-viral effect is indicatedby a reduction in cytokine levels and/or NS1 protein levels.

Typically, reductions in viremia of about 5-100 fold, more typically10-60 fold, most typically 20-30 fold, are obtained with 5-50 mg/kg biddosages of the compounds of the invention.

HCV Assay Protocol

The anti-HCV activity of the compounds of this invention was tested in ahuman hepatoma Huh-7 cell line harboring a HCV replicon. The assaycomprised the following steps:

Step 1: Compound Preparation and Serial Dilution.

Serial dilution was performed in 100% DMSO in a 384-well plate. Asolution containing a compound at 225-fold concentration of the startingfinal serial dilution concentration was prepared in 100% DMSO and 15 uLadded to the pre-specified wells in column 3 or 13 of a polypropylene384-well plate. The rest of the 384-well plate was filled with 10 uL100% DMSO except for columns 23 and 24, where 10 uL of 500 uM a HCVprotease inhibitor (ITMN-191) in 100% DMSO was added. The HCV proteaseinhibitor was used a control of 100% inhibition of HCV replication. Theplate was then placed on a Biomek FX Workstation to start the serialdilution. The serial dilution was performed for ten cycles of 3-folddilution from column 3 to 12 or from column 13 to 22.

Step 2: Cell Culture Plate Preparation and Compound Addition

To each well of a black polypropylene 384-well plate, 90 μL of cellmedia containing 1600 suspended Huh-7 HCV replicon cells was added witha Biotek uFlow Workstation. A volume of 0.4 μL of the compound solutionwas transferred from the serial dilution plate to the cell culture plateon a Biomek FX Workstation. The DMSO concentration in the final assaycondition was 0.44%. The plates were incubated for 3 days at 37° C. with5% CO2 and 85% humidity.

Step 3: Detection of Cytotoxicity and Inhibition of Viral Replication

a) Assessment of cytotoxicity: The media in the 384-well cell cultureplate was aspirated with a Biotek EL405 plate-washer. A volume of 50 μLof a solution containing 400 nM Calcein AM in 100% PBS was added to eachwell of the plate with a Biotek uFlow Workstation. The plate wasincubated for 30 minutes at room temperature before the fluorescencesignal (emission 490 nm, exitation 520 nm) was measured with a PerkinElmer Envision Plate Reader.

b) Assessment of inhibition of viral replication: The calcein-PBSsolution in the 384-well cell culture plate was aspirated with a BiotekEL405 plate-washer. A volume of 20 μL of Dual-Glo luciferase buffer(Promega, Dual-Glo Luciferase Assay Reagent, cat. #E298B) was added toeach well of the plate with a Biotek uFlow Workstation. The plate wasincubated for 10 minutes at room temperature. A volume of 20 μL of asolution containing 1:100 mixture of Dual-Glo Stop & Glosubstrate(Promega, Dual-Glo Luciferase Assay Reagent, cat. #E313B) andDual-Glo Stop & Glo buffer (Promega, Dual-Glo Luciferase Assay Reagent,cat. #E314B) was then added to each well of the plate with a BiotekuFlow Workstation. The plate was incubated at room temperature for 10minutes before the luminescence signal was measured with a Perkin ElmerEnvision Plate Reader.

Step 4: Calculation

The percent cytotoxicity was determined by calcein AM conversion tofluorescent product. The average fluorescent signal from the DMSOcontrol wells were defined as 100% nontoxic. The individual fluorescentsignal from testing compound treated well was divided by the averagesignal from DMSO control wells and then multiplied by 100% to get thepercent viability. The percent anti-HCV replication activity wasdetermined by the luminescence signal from the testing well compared toDMSO controls wells. The background signal was determined by the averageluminescence signal from the HCV protease inhibitor treated wells andwas subtracted from the signal from the testing wells as well as theDMSO control wells. Following 3-fold serial dilutions, the EC₅₀ and CC₅₀values were calculated by fitting % inhibition at each concentration tothe following equation:% inhibition=100%/[(EC ₅₀ /[I])^(b)+1]

Where b is Hill's coefficient. See, for reference, Hill, A. V., ThePossible Effects of the Aggregation of the Molecules of Haemoglobin onits Dissociation Curves, J. Physiol. 40: iv-vii. (1910).

% inhibition values at a specific concentration, for example 2 μM, canalso be derived from the formula above.

When tested, certain compounds of this invention were found to inhibitviral replication as listed in Table 1:

TABLE 1 Compound No. % inhibition at 2 μM 1 99.8 2 99.8 3 99.6 4 94.7 5100.0 6 93.2 7 99.8 8 99.8 9 99.9 10 100.0 11 99.8 12 98.2 13 99.8 1499.8 15 98.9 16 99.5 17 99.5 18 98.8 19 94.2 20 56.3 21 79.2 22 12.3

Preferred compounds of the invention have an EC₅₀ of 500 nM or less.Non-limiting examples of preferred compounds are compounds 1, 2, 3, 5,7, 8, 9, 10, 11, 13, 14, 15, 16, 17, 18, 19, 23 and 24.

The specific pharmacological and biochemical responses observed may varyaccording to and depending on the particular active compound selected orwhether there are present pharmaceutical carriers, as well as the typeof formulation and mode of administration employed, and such expectedvariations or differences in the results are contemplated in accordancewith practice of the present invention.

Although specific embodiments of the present invention are hereinillustrated and described in detail, the invention is not limitedthereto. The above detailed descriptions are provided as exemplary ofthe present invention and should not be construed as constituting anylimitation of the invention. Modifications will be obvious to thoseskilled in the art, and all modifications that do not depart from thespirit of the invention are intended to be included with the scope ofthe appended claims.

What is claimed is:
 1. A compound of Formula I:

or a pharmaceutically acceptable salt or ester thereof, wherein: R¹ is-(L¹)_(m)-(L²)_(n)-X¹; L¹ is selected from the group consisting ofalkynylene and optionally substituted arylene; m is 1; when L¹ issubstituted, L¹ is substituted with one or more Q⁶; L² is selected fromthe group consisting of —NHC(O)— and optionally substituted alkylene; nis 0 or 1; when L² is substituted, L² is substituted with one or moreQ⁶; X¹ is selected from the group consisting of a)—P(O)R^(x)R^(y), b)—P(O)OR^(x)R^(y), c)—OP(O)R^(x)R^(y) and d) optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂ cycloalkyl, optionallysubstituted C₆₋₁₄ aryl, optionally substituted 3-14 membered heteroaryl,optionally substituted 3-12 membered heterocyclyl, optionallysubstituted 3-18 membered heteroarylalkyl and optionally substitutedC₆₋₁₈ arylalkyl; each of R^(x) and R^(y) independently is selected fromthe group consisting of H, optionally substituted C₁₋₁₂ alkyl,optionally substituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂alkynyl, optionally substituted C₃₋₁₂ cycloalkyl, optionally substitutedC₆₋₁₄ aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; orR^(x) and R^(y) taken together with the atoms to which they are attachedform a 3 to 10 membered heterocyclyl; when either R^(x) or R^(y) issubstituted, R^(x) or R^(y) is substituted with one or more Q⁶; when X¹is selected from d) and is substituted, X¹ is substituted with one ormore Q¹; each Q¹ individually is selected from the group consisting ofhalogen, oxo, oxide, —NO₂, —SR¹¹, —S(O)R¹⁰, —S(O)₂R¹⁰, —S(O)₂NR¹²R¹¹,—NR¹²C(O)R¹¹, —NR¹²C(O)NR¹¹R¹², —NR¹¹S(O)R¹⁰, —NR¹¹S(O)₂R¹⁰,—NR¹²S(O)₂NR¹¹R¹², —CR¹²(═NNR¹¹R¹²), —CR¹²(═NOR¹¹), —ONR¹²R¹¹,—ON(═CR¹²R¹¹), —NR¹²OR¹¹, —OH, —NR¹¹R¹², —C(O)OR¹², —CN, —N₃,—C(═NR¹³)NR¹¹R¹², —NR¹²C(═NR¹³)NR¹¹R¹², —NR¹²C(O)OR¹¹, —OC(O)NR¹¹R¹²,—OP(O)R¹¹R¹², —P(O)R¹¹R¹², —P(O)OR¹¹R¹², —C(O)NR¹¹R¹², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(O)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted-3-10 membered heterocyclyl; whereineach R¹⁰, independently, is selected from the group consisting ofoptionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; wherein eachR¹¹ and R¹², independently, is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; or R¹¹ andR¹² taken together with the atoms to which they are attached form a 4 to10 membered heterocyclyl; or R¹⁰ and R¹¹ taken together with the atomsto which they are attached form a 4 to 10 membered heterocyclyl; eachR¹³ independently is selected from the group consisting of H, optionallysubstituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂ alkenyl,optionally substituted C₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionally substituted3-14 membered heteroaryl, optionally substituted 3-12 memberedheterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,optionally substituted C₆₋₁₈ arylalkyl, —CN, —C(O)R¹⁴, —CHO and—S(O)₂R¹⁴; where each R¹⁴ individually is optionally substituted C₁₋₁₂alkyl; when Q¹ is substituted, Q¹ is substituted with one or more Q⁶; R²is selected from the group consisting of optionally substituted C₁₋₁₂alkyl, optionally substituted C₂₋₁₂ alkenyl, optionally substitutedC₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂ cycloalkyl, optionallysubstituted C₆₋₁₄ aryl, optionally substituted 3-14 membered heteroaryl,optionally substituted 3-12 membered heterocyclyl, optionallysubstituted 3-18 membered heteroarylalkyl and optionally substitutedC₆₋₁₈ arylalkyl; wherein, when R² is substituted, R² is substituted withone or more Q²; where each Q² individually is selected from the groupconsisting of halogen, oxo, oxide, —NO₂, —SR²¹, —S(O)R²⁰, —S(O)₂R²⁰,—S(O)₂NR²²R²¹, —NR²²C(O)R²¹, —NR²²C(O)NR²¹R²², —NR²¹S(O)R²⁰,—NR²¹S(O)₂R²⁰, —NR²²S(O)₂NR²¹R²², —CR²²(═NNR²¹R²²), —CR²²(═NOR²¹),—ONR²²R²¹, —ON(═CR²²R²¹), —NR²²OR²¹, —OH, —NR²¹R²², —C(O)OR²², —CN, —N₃,—C(═NR²³)NR²¹R²², —NR²²C(═NR²³)NR²¹R²², —NR²²C(O)OR²¹, —OC(O)NR²¹R²²,—OP(O)R²¹R²², —P(O)R²¹R²², P(O)OR²¹R²², —C(O)NR²¹R²², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(O)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted 3-10 membered heterocyclyl; whereineach R²⁰, independently, is selected from the group consisting ofoptionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; wherein eachR²¹ and R²², independently, is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; or R²¹ andR²² taken together with the atoms to which they are attached form a 4 to10 membered heterocyclyl; or R²⁰ and R²¹ taken together with the atomsto which they are attached form a 4 to 10 membered heterocyclyl; eachR²³ independently is selected from the group consisting of H, optionallysubstituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂ alkenyl,optionally substituted C₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionally substituted3-14 membered heteroaryl, optionally substituted 3-12 memberedheterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,optionally substituted C₆₋₁₈ arylalkyl, —CN, —C(O)R²⁴, CHO and—S(O)₂R²⁴; where each R²⁴ individually is optionally substituted C₁₋₁₂alkyl; when Q² is substituted, Q² is substituted with one or more Q⁶; R³is -(L³)_(p)-(L⁴)_(q)-(X³); L³ is selected from the group consisting ofoptionally substituted C₁₋₁₂ alkylene, optionally substituted C₂₋₁₂alkenylene, optionally substituted C₂₋₁₂ alkynylene, optionallysubstituted C₃₋₁₂ cycloalkylene, optionally substituted C₁₋₁₂haloalkylene, optionally substituted C₆₋₁₄ arylene, optionallysubstituted 3-12 membered heterocyclylene, optionally substituted 3-18membered heteroarylalkylene and optionally substituted C₆₋₁₈arylalkylene; p is 0 or 1; when L³ is substituted, L³ is substitutedwith one or more Q³; each Q³ individually is selected from the groupconsisting of halogen, oxo, oxide, —NO₂, —SR³¹, —S(O)R³⁰, —S(O)₂R³⁰,—S(O)₂NR³²R³¹, —NR³²C(O)R³¹, —NR³²C(O)NR³¹R³², —NR³¹S(O)R³⁰,—NR³¹S(O)₂R³⁰, —NR³²S(O)₂NR³¹R³², —CR³²(═NNR³¹R³²), —CR³²(═NOR³¹),—ONR³²R³¹, —ON(═CR³²R³¹), —NR³²OR³¹, —OH, —NR³¹R³², —C(O)OR³², —CN, —N₃,—C(═NR³³)NR³¹R³², —NR³⁰C(═NR³³)N R³¹R³², —NR³⁰C(O)OR³¹, —OC(O)NR³¹R³²,—OP(O)R³¹R³², —P(O)R³¹R³², —P(O)OR³¹R³²—C(O)NR³¹R³², optionallysubstituted C₁₋₆ alkyl, optionally substituted C₂₋₆ alkenyl, optionallysubstituted C₂₋₆ alkynyl, optionally substituted C₃₋₆ cycloalkyl,optionally substituted C₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted C₁₋₆ alkyloxy, optionally substituted C₂₋₆ alkenyloxy,optionally substituted C₂₋₆ alkynyloxy, optionally substituted C₃₋₆cycloalkyloxy, optionally substituted C₆₋₁₂ aryloxy, optionallysubstituted 3-14 membered heteroaryloxy, optionally substituted 4-12membered heterocyclyloxy, optionally substituted —C(O)C₁₋₆ alkyl,optionally substituted —C(O)C₂₋₆ alkenyl, optionally substituted—C(O)C₂₋₆ alkynyl, optionally substituted —C(O)C₃₋₆ cycloalkyl,optionally substituted —C(O)C₆₋₁₂ aryl, optionally substituted—C(O)-3-14 membered heteroaryl, optionally substituted —C(O)C₆₋₁₂arylalkyl and optionally substituted 3-10 membered heterocyclyl; whereineach R³⁰, independently, is selected from the group consisting ofoptionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; wherein eachR³¹ and R³², independently, is selected from the group consisting of H,optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; or R³¹ andR³² taken together with the atoms to which they are attached form a 4 to10 membered heterocyclyl; or R³⁰ and R³¹ taken together with the atomsto which they are attached form a 4 to 10 membered heterocyclyl; eachR³³ independently is selected from the group consisting of H, optionallysubstituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂ alkenyl,optionally substituted C₂₋₁₂ alkynyl, optionally substituted C₃₋₁₂cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionally substituted3-14 membered heteroaryl, optionally substituted 3-12 memberedheterocyclyl, optionally substituted 3-18 membered heteroarylalkyl,optionally substituted C₆₋₁₈ arylalkyl, —CN, —C(O)R³⁴, CHO and—S(O)₂R³⁴; where each R³⁴ individually is optionally substituted C₁₋₁₂alkyl; when Q³ is substituted, Q³ is substituted with one or more Q⁶; L⁴is selected from the group consisting of —NH—, and optionallysubstituted C₁₋₁₂ alkylene; q is 0, 1 or 2; when L⁴ is substituted, L⁴is substituted with one or more Q⁶; X³ is selected from the groupconsisting of: e) —P(O)R^(x)R^(y), f) —P(O)OR^(x)R^(y),g)—OP(O)R^(x)R^(y), h) —P(O)(OR^(x))(OR^(y)), i)

wherein each A is a 4 to 6 membered ring, including the depicted P atom,j)

 and k) optionally substituted C₁₋₁₂ alkyl, optionally substituted C₂₋₁₂alkenyl, optionally substituted C₂₋₁₂ alkynyl, optionally substitutedC₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted 3-12membered heterocyclyl, optionally substituted 3-18 memberedheteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; each ofR^(x) and R^(y) individually is as defined above; when X³ is selectedfrom k) and is substituted, X³ is substituted with one or more Q⁴; eachQ⁴ individually is selected from the group consisting of halogen, oxo,oxide, —NO₂, —SR⁴¹, —S(O)R⁴⁰, —S(O)₂R⁴⁰, —S(O)₂NR⁴²R⁴¹, —NR⁴²C(O)R⁴¹,—NR⁴²C(O)NR⁴¹R⁴², —NR⁴¹S(O)R⁴⁰, —NR⁴¹S(O)₂R⁴⁰, —NR⁴²S(O)₂NR⁴¹R⁴²,—CR⁴²(═NNR⁴¹R⁴²), —CR⁴²(═NOR⁴¹), —ONR⁴²R⁴¹, —ON(═CR⁴²R⁴¹), —NR⁴²OR⁴¹,—OH, —NR⁴¹R⁴², —C(O)OR⁴², —CN, —N₃, —C(═NR⁴³)NR⁴¹R⁴²,—NR⁴²C(═NR⁴³)NR⁴¹R⁴², —NR⁴²C(O)OR⁴¹, and —OC(O)NR⁴¹R⁴², —OP(O)R⁴¹R⁴²,—P(O)R⁴¹R⁴², —P(O)OR⁴¹R⁴², —C(O)NR⁴¹R⁴², optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆ alkenyl, optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₆ cycloalkyl, optionally substitutedC₆₋₁₂ arylalkyl, optionally substituted C₆₋₁₂ aryl, optionallysubstituted 3-14 membered heteroaryl, optionally substituted C₁₋₆alkyloxy, optionally substituted C₂₋₆ alkenyloxy, optionally substitutedC₂₋₆ alkynyloxy, optionally substituted C₃₋₆ cycloalkyloxy, optionallysubstituted C₆₋₁₂ aryloxy, optionally substituted 3-14 memberedheteroaryloxy, optionally substituted 4-12 membered heterocyclyloxy,optionally substituted —C(O)C₁₋₆ alkyl, optionally substituted —C(O)C₂₋₆alkenyl, optionally substituted —C(O)C₂₋₆ alkynyl, optionallysubstituted —C(O)C₃₋₆ cycloalkyl, optionally substituted —C(O)C₆₋₁₂aryl, optionally substituted —C(O)-3-14 membered heteroaryl, optionallysubstituted —C(O)C₆₋₁₂ arylalkyl and optionally substituted-3-10membered heterocyclyl; wherein each R⁴⁰, independently, is selected fromthe group consisting of optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl;wherein each R⁴¹ and R⁴², independently, is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl and optionally substituted C₆₋₁₈ arylalkyl; orR⁴¹ and R⁴² taken together with the atoms to which they are attachedform a 4 to 10 membered heterocyclyl; or R⁴⁰ and R⁴¹ taken together withthe atoms to which they are attached form a 4 to 10 memberedheterocyclyl; each R⁴³ independently is selected from the groupconsisting of H, optionally substituted C₁₋₁₂ alkyl, optionallysubstituted C₂₋₁₂ alkenyl, optionally substituted C₂₋₁₂ alkynyl,optionally substituted C₃₋₁₂ cycloalkyl, optionally substituted C₆₋₁₄aryl, optionally substituted 3-14 membered heteroaryl, optionallysubstituted 3-12 membered heterocyclyl, optionally substituted 3-18membered heteroarylalkyl, optionally substituted C₆₋₁₈ arylalkyl, —CN,—C(O)R⁴⁴, CHO and —S(O)₂R⁴⁴; where each R⁴⁴ individually is optionallysubstituted C₁₋₁₂ alkyl; when Q⁴ is substituted, Q⁴ is substituted withone or more Q⁶; and each Q⁶ individually is selected from the groupconsisting of halogen, oxo, oxide, —NO₂, —N(═O), —SR⁶¹, —S(O)R⁶⁰,—S(O)₂R⁶⁰, —S(O)₂NR⁶²R⁶¹, —NR⁶²C(O)R⁶¹, —NR⁶²C(O)NR⁶¹R⁶², —NR⁶¹S(O)R⁶⁰,—NR⁶¹S(O)₂R⁶⁰, —NR⁶²S(O)₂NR⁶¹R⁶², —CR⁶²(═NNR⁶¹R⁶²), —CR⁶²(═NOR⁶¹),—ONR⁶²R⁶¹, —ON(═CR⁶²R⁶¹), —NR⁶²OR⁶¹, —OH, —NR⁶¹R⁶², —C(O)OR⁶², —CN, —N₃,—C(═NR⁶³)NR⁶¹R⁶², —NR⁶²C(═NR⁶³)NR⁶¹R⁶², —NR⁶²C(O)OR⁶¹, —OC(O)NR⁶¹R⁶²,—OP(O)R⁶¹R⁶², —P(O)R⁶¹R⁶², —P(O)OR⁶¹R⁶², —P(O)(OR⁶¹)OR⁶², —C(O)NR⁶¹R⁶²,C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, C₆₋₁₂arylalkyl, C₆₋₁₂ aryl, 3-14 membered heteroaryl, C₁₋₆ alkyloxy, C₂₋₆alkenyloxy, C₂₋₆ alkynyloxy, C₃₋₆ cycloalkyloxy, C₆₋₁₂ aryloxy, 3-14membered heteroaryloxy, 4-12 membered heterocyclyloxy, —C(O)C₁₋₆ alkyl,—C(O)C₂₋₆ alkenyl, —C(O)C₂₋₆ alkynyl, —C(O)C₃₋₆ cycloalkyl, —C(O)C₁₋₆haloalkyl, —C(O)C₆₋₁₂ aryl, —C(O)-3-14 membered heteroaryl, —C(O)C₆₋₁₂arylalkyl and 3-10 membered heterocyclyl; wherein each R⁶⁰,independently, is selected from the group consisting of C₁₋₁₂ alkyl,C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂ cycloalkyl, C₁₋₁₂ haloalkyl, C₆₋₁₄aryl, 3-14 membered heteroaryl, 3-12 membered heterocyclyl, 3-18membered heteroarylalkyl and C₆₋₁₈ arylalkyl; wherein each R⁶¹ and R⁶²,independently, is selected from the group consisting of H, C₁₋₁₂ alkyl,C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂ cycloalkyl, C₁₋₁₂ haloalkyl, C₆₋₁₄aryl, 3-14 membered heteroaryl, 3-12 membered heterocyclyl, 3-18membered heteroarylalkyl and C₆₋₁₈ arylalkyl; or R⁶¹ and R⁶² takentogether with the atoms to which they are attached form a 4 to 10membered heterocyclyl; or R⁶⁰ and R⁶¹ taken together with the atoms towhich they are attached form a 4 to 10 membered heterocyclyl; each R⁶³independently is selected from the group consisting of H, C₁₋₁₂ alkyl,C₂₋₁₂ alkenyl, C₂₋₁₂ alkynyl, C₃₋₁₂ cycloalkyl, C₆₋₁₄ aryl, 3-14membered heteroaryl, 3-12 membered heterocyclyl, 3-18 memberedheteroarylalkyl, C₆₋₁₈ arylalkyl, —CN, —C(O)R⁶⁴, —CHO and —S(O)₂R⁶⁴;where each R⁶⁴ individually is C₁₋₁₂ alkyl; provided, however, that whenX¹ is selected from d), then X³ is selected from the group consisting ofe), f), g), h), i) and j).
 2. The compound of claim 1, wherein L¹ isphenylene.
 3. The compound of claim 1, wherein L¹ is —C≡C—; n is 0; andX¹ is optionally substituted C₁₋₁₂ alkyl.
 4. The compound of claim 1,wherein the compound of Formula I is represented by a compound ofFormula II:

or a pharmaceutically acceptable salt or ester thereof.
 5. The compoundof claim 1, wherein R² is optionally substituted C₃₋₁₂ cycloalkyl. 6.The compound of claim 5, wherein R² is optionally substitutedcyclohexyl.
 7. The compound of claim 1, wherein R² is selected from thegroup consisting of:


8. The compound of claim 7, wherein R² is


9. The compound of claim 1, wherein p is
 1. 10. The compound of claim 1,wherein p is
 0. 11. The compound of claim 1, wherein L³ is optionallysubstituted C₃₋₁₂ cycloalkylene.
 12. The compound of claim 11, whereinL³ is optionally substituted cyclohexylene.
 13. The compound of claim 1,where L³ is an optionally substituted 5-6 membered N-containingheterocyclene.
 14. The compound of claim 1, wherein L³ is optionallysubstituted C₁₋₁₂ alkylene.
 15. The compound of claim 1, wherein q is 0.16. The compound of claim 1, wherein q is
 1. 17. The compound of claim1, wherein q is
 2. 18. The compound of claim 1, wherein X³ is


19. The compound of claim 1, wherein X³ is —P(O)R^(x)R^(y).
 20. Thecompound of claim 1, wherein X³ is —P(O)OR^(x)R^(y).
 21. The compound ofclaim 1, wherein X³ is —OP(O)R^(x)R^(y).
 22. The compound of claim 1,wherein X³ is —P(O)(OR^(x))(OR^(y)).
 23. The compound of claim 1selected from the group consisting of

or a pharmaceutically acceptable salt or ester thereof.
 24. Apharmaceutical composition comprising a therapeutically effective amountof a compound of claim 1 and a pharmaceutically acceptable carrier orexcipient.
 25. The pharmaceutical composition of claim 24 furthercomprising at least one therapeutic agent selected from the groupconsisting of an interferon, ribavirin or an analog thereof, an HCV NS3protease inhibitor, an NS5a inhibitor, an alpha-glucosidase 1 inhibitor,a hepatoprotectant, a mevalonate decarboxylase antagonist, an antagonistof the renin-angiotensin system, an endothelin antagonist, otheranti-fibrotic agents, a nucleoside or nucleotide inhibitor of HCV NS5Bpolymerase, a non-nucleoside inhibitor of HCV NS5B polymerase, an HCVNS5A inhibitor, a TLR-7 agonist, a cyclophillin inhibitor, an HCV IRESinhibitor, a pharmacokinetic enhancer and other drugs for treating HCV;or a mixture thereof.
 26. A method for treating a Flaviviridae viralinfection comprising administering a therapeutically effective amount ofa compound of claim 1 to a mammal in need thereof.
 27. The method ofclaim 26 wherein the viral infection is caused by a Hepatitis C virus.28. The method of claim 27 further comprising administering at least onetherapeutic agent selected from the group consisting of an interferon,ribavirin or an analog thereof, an HCV NS3 protease inhibitor, an NS5ainhibitor, an alpha-glucosidase 1 inhibitor, a hepatoprotectant, amevalonate decarboxylase antagonist, an antagonist of therenin-angiotensin system, an endothelin antagonist, other anti-fibroticagent, a nucleoside or nucleotide inhibitor of HCV NS5B polymerase, anon-nucleoside inhibitor of HCV NS5B polymerase, an HCV NS5A inhibitor,a TLR-7 agonist, a cyclophillin inhibitor, an HCV IRES inhibitor, apharmacokinetic enhancer and other drugs for treating HCV; or a mixturethereof.